Cell culture

Cell culture or Tissue culture is the process by which cells are grown under controlled conditions, generally outside their natural environment. The term "tissue culture" was coined by American pathologist Montrose Thomas Burrows.[1] This technique is also called micropropagation. After the cells of interest have been isolated from living tissue, they can subsequently be maintained under carefully controlled conditions the need to be kept at body temperature (37 °C) in an incubator.[2] These conditions vary for each cell type, but generally consist of a suitable vessel with a substrate or rich medium that supplies the essential nutrients (amino acids, carbohydrates, vitamins, minerals), growth factors, hormones, and gases (CO2, O2), and regulates the physio-chemical environment (pH buffer, osmotic pressure, temperature). Most cells require a surface or an artificial substrate to form an adherent culture as a monolayer (one single-cell thick), whereas others can be grown free floating in a medium as a suspension culture.[3] This is typically facilitated via use of a liquid, semi-solid, or solid growth medium, such as broth or agar. Tissue culture commonly refers to the culture of animal cells and tissues, with the more specific term plant tissue culture being used for plants. The lifespan of most cells is genetically determined, but some cell culturing cells have been “transformed” into immortal cells which will reproduce indefinitely if the optimal conditions are provided.

"Co-culture" redirects here. For the concept of cultures-within-cultures, see Subculture.
Cell culture in a small Petri dish
Epithelial cells in culture, stained for keratin (red) and DNA (green)

In practice, the term "cell culture" now refers to the culturing of cells derived from multicellular eukaryotes, especially animal cells, in contrast with other types of culture that also grow cells, such as plant tissue culture, fungal culture, and microbiological culture (of microbes). The historical development and methods of cell culture are closely interrelated to those of tissue culture and organ culture. Viral culture is also related, with cells as hosts for the viruses.

The laboratory technique of maintaining live cell lines (a population of cells descended from a single cell and containing the same genetic makeup) separated from their original tissue source became more robust in the middle 20th century.[4][5]

History

The 19th-century English physiologist Sydney Ringer developed salt solutions containing the chlorides of sodium, potassium, calcium and magnesium suitable for maintaining the beating of an isolated animal heart outside the body.[6] In 1885 Wilhelm Roux removed a section of the medullary plate of an embryonic chicken and maintained it in a warm saline solution for several days, establishing the basic principle of tissue culture. In 1907 the zoologist Ross Granville Harrison demonstrated the growth of frog embryonic cells that would give rise to nerve cells in a medium of clotted lymph. In 1913, E. Steinhardt, C. Israeli, and R. A. Lambert grew vaccinia virus in fragments of guinea pig corneal tissue.[7] In 1996, the first use of regenerative tissue was used to replace a small length of urethra, which led to the understanding that the technique of obtaining samples of tissue, growing it outside the body without a scaffold, and reapplying it, can be used for only small distances of less than 1 cm.[8][9][10] Ross Granville Harrison, working at Johns Hopkins Medical School and then at Yale University, published results of his experiments from 1907 to 1910, establishing the methodology of tissue culture.[11]

Gottlieb Haberlandt first pointed out the possibilities of the culture of isolated tissues, plant tissue culture.[12] He suggested that the potentialities of individual cells via tissue culture as well as that the reciprocal influences of tissues on one another could be determined by this method. Since Haberlandt's original assertions, methods for tissue and cell culture have been realized, leading to significant discoveries in biology and medicine. His original idea, presented in 1902, was called totipotentiality: “Theoretically all plant cells are able to give rise to a complete plant.”[13][14][15]

Cell culture techniques were advanced significantly in the 1940s and 1950s to support research in virology. Growing viruses in cell cultures allowed preparation of purified viruses for the manufacture of vaccines. The injectable polio vaccine developed by Jonas Salk was one of the first products mass-produced using cell culture techniques. This vaccine was made possible by the cell culture research of John Franklin Enders, Thomas Huckle Weller, and Frederick Chapman Robbins, who were awarded a Nobel Prize for their discovery of a method of growing the virus in monkey kidney cell cultures. Cell culture has contributed to the development of vaccines for many diseases.[2]

Modern usage
Cultured cells growing in growth medium

In modern usage, "tissue culture" generally refers to the growth of cells from a tissue from a multicellular organism in vitro. These cells may be cells isolated from a donor organism (primary cells) or an immortalised cell line. The cells are bathed in a culture medium, which contains essential nutrients and energy sources necessary for the cells' survival.[16] Thus, in its broader sense, "tissue culture" is often used interchangeably with "cell culture". On the other hand, the strict meaning of "tissue culture" refers to the culturing of tissue pieces, i.e. explant culture.

Tissue culture is an important tool for the study of the biology of cells from multicellular organisms. It provides an in vitro model of the tissue in a well defined environment which can be easily manipulated and analysed. In animal tissue culture, cells may be grown as two-dimensional monolayers (conventional culture) or within fibrous scaffolds or gels to attain more naturalistic three-dimensional tissue-like structures (3D culture). Eric Simon, in a 1988 NIH SBIR grant report, showed that electrospinning could be used to produced nano- and submicron-scale polymeric fibrous scaffolds specifically intended for use as in vitro cell and tissue substrates. This early use of electrospun fibrous lattices for cell culture and tissue engineering showed that various cell types would adhere to and proliferate upon polycarbonate fibers. It was noted that as opposed to the flattened morphology typically seen in 2D culture, cells grown on the electrospun fibers exhibited a more rounded 3-dimensional morphology generally observed of tissues in vivo.[17]

Plant tissue culture in particular is concerned with the growing of entire plants from small pieces of plant tissue, cultured in medium.[18]

Concepts in mammalian cell culture

Isolation of cells

Cells can be isolated from tissues for ex vivo culture in several ways. Cells can be easily purified from blood; however, only the white cells are capable of growth in culture. Cells can be isolated from solid tissues by digesting the extracellular matrix using enzymes such as collagenase, trypsin, or pronase, before agitating the tissue to release the cells into suspension.[19][20] Alternatively, pieces of tissue can be placed in growth media, and the cells that grow out are available for culture. This method is known as explant culture.

Cells that are cultured directly from a subject are known as primary cells. With the exception of some derived from tumors, most primary cell cultures have limited lifespan.

An established or immortalized cell line has acquired the ability to proliferate indefinitely either through random mutation or deliberate modification, such as artificial expression of the telomerase gene. Numerous cell lines are well established as representative of particular cell types.

Maintaining cells in culture

For the majority of isolated primary cells, they undergo the process of senescence and stop dividing after a certain number of population doublings while generally retaining their viability (described as the Hayflick limit).

A bottle of DMEM cell culture medium

Aside from temperature and gas mixture, the most commonly varied factor in culture systems is the cell growth medium. Recipes for growth media can vary in pH, glucose concentration, growth factors, and the presence of other nutrients. The growth factors used to supplement media are often derived from the serum of animal blood, such as fetal bovine serum (FBS), bovine calf serum, equine serum, and porcine serum. One complication of these blood-derived ingredients is the potential for contamination of the culture with viruses or prions, particularly in medical biotechnology applications. Current practice is to minimize or eliminate the use of these ingredients wherever possible and use human platelet lysate (hPL).[21] This eliminates the worry of cross-species contamination when using FBS with human cells. hPL has emerged as a safe and reliable alternative as a direct replacement for FBS or other animal serum. In addition, chemically defined media can be used to eliminate any serum trace (human or animal), but this cannot always be accomplished with different cell types. Alternative strategies involve sourcing the animal blood from countries with minimum BSE/TSE risk, such as The United States, Australia and New Zealand,[22] and using purified nutrient concentrates derived from serum in place of whole animal serum for cell culture.[23]

Plating density (number of cells per volume of culture medium) plays a critical role for some cell types. For example, a lower plating density makes granulosa cells exhibit estrogen production, while a higher plating density makes them appear as progesterone-producing theca lutein cells.[24]

Cells can be grown either in suspension or adherent cultures.[25] Some cells naturally live in suspension, without being attached to a surface, such as cells that exist in the bloodstream. There are also cell lines that have been modified to be able to survive in suspension cultures so they can be grown to a higher density than adherent conditions would allow. Adherent cells require a surface, such as tissue culture plastic or microcarrier, which may be coated with extracellular matrix (such as collagen and laminin) components to increase adhesion properties and provide other signals needed for growth and differentiation. Most cells derived from solid tissues are adherent. Another type of adherent culture is organotypic culture, which involves growing cells in a three-dimensional (3-D) environment as opposed to two-dimensional culture dishes. This 3D culture system is biochemically and physiologically more similar to in vivo tissue, but is technically challenging to maintain because of many factors (e.g. diffusion).[26]

Cell culture basal media

There are different kinds of cell culture media which being used routinely in life science including the following:

  • MEM
  • DMEM
  • RPMI 1640
  • Ham's f-12
  • IMDM
  • Leibovitz L-15
  • DMEM/F-12

Components of cell culture media
Component Function
Carbon source (glucose/glutamine) Source of energy
Amino acid Building blocks of protein
Vitamins Promote cell survival and growth
Balanced salt solution An isotonic mixture of ions to maintain optimum osmotic pressure within the cells and provide essential metal ions to act as cofactors for enzymatic reactions, cell adhesion etc.
Phenol red dye pH indicator. The color of phenol red changes from orange/red at pH 7–7.4 to yellow at acidic (lower) pH and purple at basic (higher) pH.
Bicarbonate /HEPES buffer It is used to maintain a balanced pH in the media

Typical Growth conditions
Parameter
Temperature 37  °C
CO2 5%
Relative Humidity 95%

Cell line cross-contamination

Cell line cross-contamination can be a problem for scientists working with cultured cells.[27] Studies suggest anywhere from 15 to 20% of the time, cells used in experiments have been misidentified or contaminated with another cell line.[28][29][30] Problems with cell line cross-contamination have even been detected in lines from the NCI-60 panel, which are used routinely for drug-screening studies.[31][32] Major cell line repositories, including the American Type Culture Collection (ATCC), the European Collection of Cell Cultures (ECACC) and the German Collection of Microorganisms and Cell Cultures (DSMZ), have received cell line submissions from researchers that were misidentified by them.[31][33] Such contamination poses a problem for the quality of research produced using cell culture lines, and the major repositories are now authenticating all cell line submissions.[34] ATCC uses short tandem repeat (STR) DNA fingerprinting to authenticate its cell lines.[35]

To address this problem of cell line cross-contamination, researchers are encouraged to authenticate their cell lines at an early passage to establish the identity of the cell line. Authentication should be repeated before freezing cell line stocks, every two months during active culturing and before any publication of research data generated using the cell lines. Many methods are used to identify cell lines, including isoenzyme analysis, human lymphocyte antigen (HLA) typing, chromosomal analysis, karyotyping, morphology and STR analysis.[35]

One significant cell-line cross contaminant is the immortal HeLa cell line. Hela contamination was first noted in the early 1960s in non-human culture in the USA. Intraspecies contamination was discovered in nineteen cell lines in the seventies. In 1974, five human cell lines from the Soviet Union were found to be Hela. A follow-up study analysing 50-odd cell lines indicated that half had Hela markers, but contaminant Hela had hybridised with the original cell lines. Hela cell contamination from air droplets has been reported. Hela was even unknowingly injected into human subjects by Jonas Salk in a 1978 vaccine trial.[36]

Other technical issues

As cells generally continue to divide in culture, they generally grow to fill the available area or volume. This can generate several issues:

  • Nutrient depletion in the growth media
  • Changes in pH of the growth media
  • Accumulation of apoptotic/necrotic (dead) cells
  • Cell-to-cell contact can stimulate cell cycle arrest, causing cells to stop dividing, known as contact inhibition.
  • Cell-to-cell contact can stimulate cellular differentiation.
  • Genetic and epigenetic alterations, with a natural selection of the altered cells potentially leading to overgrowth of abnormal, culture-adapted cells with decreased differentiation and increased proliferative capacity.[37]

The choice of culture medium might affect the physiological relevance of findings from cell culture experiments due to the differences in the nutrient composition and concentrations.[38] A systematic bias in generated datasets was recently shown for CRISPR and RNAi gene silencing screens,[39] and for metabolic profiling of cancer cell lines.[38] Using a growth medium that better represents the physiological levels of nutrients can improve the physiological relevance of in vitro studies and recently such media types, as Plasmax[40] and Human Plasma Like Medium (HPLM),[41] were developed.

Manipulation of cultured cells

Among the common manipulations carried out on culture cells are media changes, passaging cells, and transfecting cells. These are generally performed using tissue culture methods that rely on aseptic technique. Aseptic technique aims to avoid contamination with bacteria, yeast, or other cell lines. Manipulations are typically carried out in a biosafety cabinet or laminar flow cabinet to exclude contaminating micro-organisms. Antibiotics (e.g. penicillin and streptomycin) and antifungals (e.g.amphotericin B and Antibiotic-Antimycotic solution) can also be added to the growth media.

As cells undergo metabolic processes, acid is produced and the pH decreases. Often, a pH indicator is added to the medium to measure nutrient depletion.

Media changes

In the case of adherent cultures, the media can be removed directly by aspiration, and then is replaced. Media changes in non-adherent cultures involve centrifuging the culture and resuspending the cells in fresh media.

Passaging cells

Passaging (also known as subculture or splitting cells) involves transferring a small number of cells into a new vessel. Cells can be cultured for a longer time if they are split regularly, as it avoids the senescence associated with prolonged high cell density. Suspension cultures are easily passaged with a small amount of culture containing a few cells diluted in a larger volume of fresh media. For adherent cultures, cells first need to be detached; this is commonly done with a mixture of trypsin-EDTA; however, other enzyme mixes are now available for this purpose. A small number of detached cells can then be used to seed a new culture. Some cell cultures, such as RAW cells are mechanically scraped from the surface of their vessel with rubber scrapers.

Transfection and transduction
Main articles: Transfection and Transformation (genetics)

Another common method for manipulating cells involves the introduction of foreign DNA by transfection. This is often performed to cause cells to express a gene of interest. More recently, the transfection of RNAi constructs have been realized as a convenient mechanism for suppressing the expression of a particular gene/protein. DNA can also be inserted into cells using viruses, in methods referred to as transduction, infection or transformation. Viruses, as parasitic agents, are well suited to introducing DNA into cells, as this is a part of their normal course of reproduction.

Established human cell lines
Cultured HeLa cells have been stained with Hoechst turning their nuclei blue, and are one of the earliest human cell lines descended from Henrietta Lacks, who died of cervical cancer from which these cells originated.

Cell lines that originate with humans have been somewhat controversial in bioethics, as they may outlive their parent organism and later be used in the discovery of lucrative medical treatments. In the pioneering decision in this area, the Supreme Court of California held in Moore v. Regents of the University of California that human patients have no property rights in cell lines derived from organs removed with their consent.[42]

It is possible to fuse normal cells with an immortalised cell line. This method is used to produce monoclonal antibodies. In brief, lymphocytes isolated from the spleen (or possibly blood) of an immunised animal are combined with an immortal myeloma cell line (B cell lineage) to produce a hybridoma which has the antibody specificity of the primary lymphocyte and the immortality of the myeloma. Selective growth medium (HA or HAT) is used to select against unfused myeloma cells; primary lymphoctyes die quickly in culture and only the fused cells survive. These are screened for production of the required antibody, generally in pools to start with and then after single cloning.

Cell strains

A cell strain is derived either from a primary culture or a cell line by the selection or cloning of cells having specific properties or characteristics which must be defined. Cell strains are cells that have been adapted to culture but, unlike cell lines, have a finite division potential. Non-immortalized cells stop dividing after 40 to 60 population doublings[43] and, after this, they lose their ability to proliferate (a genetically determined event known as senescence).[44]

Applications of cell culture

Mass culture of animal cell lines is fundamental to the manufacture of viral vaccines and other products of biotechnology. Culture of human stem cells is used to expand the number of cells and differentiate the cells into various somatic cell types for transplantation.[45] Stem cell culture is also used to harvest the molecules and exosomes that the stem cells release for the purposes of therapeutic development.[46]

Biological products produced by recombinant DNA (rDNA) technology in animal cell cultures include enzymes, synthetic hormones, immunobiologicals (monoclonal antibodies, interleukins, lymphokines), and anticancer agents. Although many simpler proteins can be produced using rDNA in bacterial cultures, more complex proteins that are glycosylated (carbohydrate-modified) currently must be made in animal cells. An important example of such a complex protein is the hormone erythropoietin. The cost of growing mammalian cell cultures is high, so research is underway to produce such complex proteins in insect cells or in higher plants, use of single embryonic cell and somatic embryos as a source for direct gene transfer via particle bombardment, transit gene expression and confocal microscopy observation is one of its applications. It also offers to confirm single cell origin of somatic embryos and the asymmetry of the first cell division, which starts the process.

Cell culture is also a key technique for cellular agriculture, which aims to provide both new products and new ways of producing existing agricultural products like milk, (cultured) meat, fragrances, and rhino horn from cells and microorganisms. It is therefore considered one means of achieving animal-free agriculture. It is also a central tool for teaching cell biology.[47]

Cell culture in two dimensions

Research in tissue engineering, stem cells and molecular biology primarily involves cultures of cells on flat plastic dishes. This technique is known as two-dimensional (2D) cell culture, and was first developed by Wilhelm Roux who, in 1885, removed a portion of the medullary plate of an embryonic chicken and maintained it in warm saline for several days on a flat glass plate. From the advance of polymer technology arose today's standard plastic dish for 2D cell culture, commonly known as the Petri dish. Julius Richard Petri, a German bacteriologist, is generally credited with this invention while working as an assistant to Robert Koch. Various researchers today also utilize culturing laboratory flasks, conicals, and even disposable bags like those used in single-use bioreactors.

Aside from Petri dishes, scientists have long been growing cells within biologically derived matrices such as collagen or fibrin, and more recently, on synthetic hydrogels such as polyacrylamide or PEG. They do this in order to elicit phenotypes that are not expressed on conventionally rigid substrates. There is growing interest in controlling matrix stiffness,[48] a concept that has led to discoveries in fields such as:

Cell culture in three dimensions

Cell culture in three dimensions has been touted as "Biology's New Dimension".[63] At present, the practice of cell culture remains based on varying combinations of single or multiple cell structures in 2D.[64] Currently, there is an increase in use of 3D cell cultures in research areas including drug discovery, cancer biology, regenerative medicine, nanomaterials assessment and basic life science research.[65][66][67] 3D cell cultures can be grown using a scaffold or matrix, or in a scaffold-free manner. Scaffold based cultures utilize an acellular 3D matrix or a liquid matrix. Scaffold-free methods are normally generated in suspensions.[68] There are a variety of platforms used to facilitate the growth of three-dimensional cellular structures including scaffold systems such as hydrogel matrices[69] and solid scaffolds, and scaffold-free systems such as low-adhesion plates, nanoparticle facilitated magnetic levitation,[70] and hanging drop plates.[71][72] Culturing cells in 3D leads to wide variation in gene expression signatures and partly mimics tissues in the physiological states.[73] A 3D cell culture model showed cell growth similar to that of in vivo than did a monolayer culture, and all three cultures were capable of sustaining cell growth.[74] As 3D culturing has been developed it turns out to have a great potential to design tumors models and investigate malignant transformation and metastasis, 3D cultures can provide aggerate tool for understanding changes, interactions, and cellular signaling.[75]

3D cell culture in scaffolds

Eric Simon, in a 1988 NIH SBIR grant report, showed that electrospinning could be used to produced nano- and submicron-scale polystyrene and polycarbonate fibrous scaffolds specifically intended for use as in vitro cell substrates. This early use of electrospun fibrous lattices for cell culture and tissue engineering showed that various cell types including Human Foreskin Fibroblasts (HFF), transformed Human Carcinoma (HEp-2), and Mink Lung Epithelium (MLE) would adhere to and proliferate upon polycarbonate fibers. It was noted that, as opposed to the flattened morphology typically seen in 2D culture, cells grown on the electrospun fibers exhibited a more histotypic rounded 3-dimensional morphology generally observed in vivo.[17]

3D cell culture in hydrogels

As the natural extracellular matrix (ECM) is important in the survival, proliferation, differentiation and migration of cells, different hydrogel culture matrices mimicking natural ECM structure are seen as potential approaches to in vivo –like cell culturing.[76] Hydrogels are composed of interconnected pores with high water retention, which enables efficient transport of substances such as nutrients and gases. Several different types of hydrogels from natural and synthetic materials are available for 3D cell culture, including animal ECM extract hydrogels, protein hydrogels, peptide hydrogels, polymer hydrogels, and wood-based nanocellulose hydrogel.

3D Cell Culturing by Magnetic Levitation

The 3D Cell Culturing by Magnetic Levitation method (MLM) is the application of growing 3D tissue by inducing cells treated with magnetic nanoparticle assemblies in spatially varying magnetic fields using neodymium magnetic drivers and promoting cell to cell interactions by levitating the cells up to the air/liquid interface of a standard petri dish. The magnetic nanoparticle assemblies consist of magnetic iron oxide nanoparticles, gold nanoparticles, and the polymer polylysine. 3D cell culturing is scalable, with the capability for culturing 500 cells to millions of cells or from single dish to high-throughput low volume systems.

Tissue culture and engineering

Cell culture is a fundamental component of tissue culture and tissue engineering, as it establishes the basics of growing and maintaining cells in vitro. The major application of human cell culture is in stem cell industry, where mesenchymal stem cells can be cultured and cryopreserved for future use. Tissue engineering potentially offers dramatic improvements in low cost medical care for hundreds of thousands of patients annually.

Vaccines

Vaccines for polio, measles, mumps, rubella, and chickenpox are currently made in cell cultures. Due to the H5N1 pandemic threat, research into using cell culture for influenza vaccines is being funded by the United States government. Novel ideas in the field include recombinant DNA-based vaccines, such as one made using human adenovirus (a common cold virus) as a vector,[77][78] and novel adjuvants.[79]

Cell Co-Culture

The technique of co-culturing is used to study cell crosstalk between two or more types of cells on a plate or in a 3D matrix. A similar in vitro model to biological tissue can be used to investigate the cultivation of different stem cells and interaction of immune cells. Since most tissues contain more than one type of cell, it is important to evaluate their interaction in a 3D culture environment to gain a better understanding of their interaction and to introduce mimetic tissues. There are two types of co-culturing: direct and indirect. Using the direct method, cells are in contact with each other in the same culture media or matrix, while using the indirect method, different environments are used, allowing the interaction to take place through signaling and soluble factors.[1][80]

The Co-Cultured System is useful for studying the model of cancer tumors, the effect of drugs during therapeutic trials, and the differentiation of cells in tissue models during interaction between cells. the coculture system in 3D model can predict the response to chemotherapy and endocrine therapy if the microenvironment defines biological tissue for the cells.

A co-culture method is used in tissue engineering to generate tissue formation with multiple cells interacting directly.[81]

Cell culture in microfluidic device

Microfluidics technique is developed systems that can perform a process in a flow which are usually in a scale of micron. Microfluidics chip are also known as Lab-on-a-chip and they are able to have continuous procedure and reaction steps with spare amount of reactants and space. Such systems enable the identification and isolation of individual cells and molecules when combined with appropriate biological assays and high-sensitivity detection techniques.[82][83]

Organ-on-a-chip

OoC systems mimic and control the microenvironment of the cells by growing tissues in microfluidics. Combining tissue engineering, biomaterials fabrication, and cell biology, it offers the possibility of establishing a biomimetic model for studying human diseases in the laboratory. In recent years, 3D cell culture science has made significant progress, leading to the development of OoC. OoC is considered as a preclinical step that benefits pharmaceutical studies, drug development and disease modeling.[84][85] OoC is an important technology that can bridge the gap between animal testing and clinical studies and also by the advances that the science has achieved could be a replace for in vivo studies for drug delivery and pathophysiological studies.[86]

Culture of non-mammalian cells

Besides the culture of well-established immortalised cell lines, cells from primary explants of a plethora of organisms can be cultured for a limited period of time before senescence occurs (see Hayflick's limit). Cultured primary cells have been extensively used in research, as is the case of fish keratocytes in cell migration studies.[87][47][88]

Plant cell culture methods

Plant cell cultures are typically grown as cell suspension cultures in a liquid medium or as callus cultures on a solid medium. The culturing of undifferentiated plant cells and calli requires the proper balance of the plant growth hormones auxin and cytokinin.

Insect cell culture

Cells derived from Drosophila melanogaster (most prominently, Schneider 2 cells) can be used for experiments which may be hard to do on live flies or larvae, such as biochemical studies or studies using siRNA. Cell lines derived from the army worm Spodoptera frugiperda, including Sf9 and Sf21, and from the cabbage looper Trichoplusia ni, High Five cells, are commonly used for expression of recombinant proteins using baculovirus.[89]

Bacterial and yeast culture methods

For bacteria and yeasts, small quantities of cells are usually grown on a solid support that contains nutrients embedded in it, usually a gel such as agar, while large-scale cultures are grown with the cells suspended in a nutrient broth.

Viral culture methods

The culture of viruses requires the culture of cells of mammalian, plant, fungal or bacterial origin as hosts for the growth and replication of the virus. Whole wild type viruses, recombinant viruses or viral products may be generated in cell types other than their natural hosts under the right conditions. Depending on the species of the virus, infection and viral replication may result in host cell lysis and formation of a viral plaque.

Common cell lines
Human cell lines
Primate cell lines
  • Vero (African green monkey Chlorocebus kidney epithelial cell line)
Mouse cell lines
  • MC3T3 (embryonic calvarium)
Rat tumor cell lines
  • GH3 (pituitary tumor)
  • PC12 (pheochromocytoma)
Plant cell lines
  • Tobacco BY-2 cells (kept as cell suspension culture, they are model system of plant cell)
Other species cell lines

List of cell lines
Cell lineMeaningOrganismOrigin tissueMorphologyLinks
3T3-L1"3-day transfer, inoculum 3 x 10^5 cells"MouseEmbryoFibroblastECACC Cellosaurus
4T1MouseMammary glandATCC Cellosaurus
1321N1HumanBrainAstrocytomaECACC Cellosaurus
9LRatBrainGlioblastomaECACC Cellosaurus
A172HumanBrainGlioblastomaECACC Cellosaurus
A20MouseB lymphomaB lymphocyteCellosaurus
A253HumanSubmandibular ductHead and neck carcinomaATCC Cellosaurus
A2780HumanOvaryOvarian carcinomaECACC Cellosaurus
A2780ADRHumanOvaryAdriamycin-resistant derivative of A2780ECACC Cellosaurus
A2780cisHumanOvaryCisplatin-resistant derivative of A2780ECACC Cellosaurus
A431HumanSkin epitheliumSquamous cell carcinomaECACC Cellosaurus
A549HumanLungLung carcinomaECACC Cellosaurus
AB9ZebrafishFinFibroblastATCC Cellosaurus
AHL-1Armenian Hamster Lung-1HamsterLungECACC Cellosaurus
ALCMouseBone marrowStromaPMID 2435412[90] Cellosaurus
B16MouseMelanomaECACC Cellosaurus
B35RatNeuroblastomaATCC Cellosaurus
BCP-1HumanPBMCHIV+ primary effusion lymphomaATCC Cellosaurus
BEAS-2BBronchial epithelium + Adenovirus 12-SV40 virus hybrid (Ad12SV40)HumanLungEpithelialECACC Cellosaurus
bEnd.3Brain Endothelial 3MouseBrain/cerebral cortexEndotheliumCellosaurus
BHK-21Baby Hamster Kidney-21HamsterKidneyFibroblastECACC Cellosaurus
BOSC23Packaging cell line derived from HEK 293HumanKidney (embryonic)EpitheliumCellosaurus
BT-20Breast Tumor-20HumanBreast epitheliumBreast carcinomaATCC Cellosaurus
BxPC-3Biopsy xenograft of Pancreatic Carcinoma line 3HumanPancreatic adenocarcinomaEpithelialECACC Cellosaurus
C2C12MouseMyoblastECACC Cellosaurus
C3H-10T1/2MouseEmbryonic mesenchymal cell lineECACC Cellosaurus
C6RatBrain astrocyteGliomaECACC Cellosaurus
C6/36Insect - Asian tiger mosquitoLarval tissueECACC Cellosaurus
Caco-2HumanColonColorectal carcinomaECACC Cellosaurus
Cal-27HumanTongueSquamous cell carcinomaATCC Cellosaurus
Calu-3HumanLungAdenocarcinomaATCC Cellosaurus
CGR8MouseEmbryonic stem cellsECACC Cellosaurus
CHOChinese Hamster OvaryHamsterOvaryEpitheliumECACC Cellosaurus
CML T1Chronic myeloid leukemia T lymphocyte 1HumanCML acute phaseT cell leukemiaDSMZ Cellosaurus
CMT12Canine Mammary Tumor 12DogMammary glandEpitheliumCellosaurus
COR-L23HumanLungLung carcinomaECACC Cellosaurus
COR-L23/5010HumanLungLung carcinomaECACC Cellosaurus
COR-L23/CPRHumanLungLung carcinomaECACC Cellosaurus
COR-L23/R23-HumanLungLung carcinomaECACC Cellosaurus
COS-7Cercopithecus aethiops, origin-defective SV-40Old World monkey - Cercopithecus aethiops (Chlorocebus)KidneyFibroblastECACC Cellosaurus
COV-434HumanOvaryOvarian granulosa cell carcinomaPMID 8436435[91] ECACC Cellosaurus
CT26MouseColonColorectal carcinomaCellosaurus
D17DogLung metastasisOsteosarcomaATCC Cellosaurus
DAOYHumanBrainMedulloblastomaATCC Cellosaurus
DH82DogHistiocytosisMonocyte/macrophageECACC Cellosaurus
DU145HumanAndrogen insensitive prostate carcinomaATCC Cellosaurus
DuCaPDura mater cancer of the ProstateHumanMetastatic prostate carcinomaEpithelialPMID 11317521[92] Cellosaurus
E14Tg2aMouseEmbryonic stem cellsECACC Cellosaurus
EL4MouseT cell leukemiaECACC Cellosaurus
EM-2HumanCML blast crisisPh+ CML lineDSMZ Cellosaurus
EM-3HumanCML blast crisisPh+ CML lineDSMZ Cellosaurus
EMT6/AR1MouseMammary glandEpithelial-likeECACC Cellosaurus
EMT6/AR10.0MouseMammary glandEpithelial-likeECACC Cellosaurus
FM3HumanLymph node metastasisMelanomaECACC Cellosaurus
GL261Glioma 261MouseBrainGliomaCellosaurus
H1299HumanLungLung carcinomaATCC Cellosaurus
HaCaTHumanSkinKeratinocyteCLS Cellosaurus
HCA2HumanColonAdenocarcinomaECACC Cellosaurus
HEK 293Human Embryonic Kidney 293HumanKidney (embryonic)EpitheliumECACC Cellosaurus
HEK 293THEK 293 derivativeHumanKidney (embryonic)EpitheliumECACC Cellosaurus
HeLa"Henrietta Lacks"HumanCervix epitheliumCervical carcinomaECACC Cellosaurus
Hepa1c1c7Clone 7 of clone 1 hepatoma line 1MouseHepatomaEpithelialECACC Cellosaurus
Hep G2HumanLiverHepatoblastomaECACC Cellosaurus
High FiveInsect (moth) - Trichoplusia niOvaryCellosaurus
HL-60Human Leukemia-60HumanBloodMyeloblastECACC Cellosaurus
HT-1080HumanFibrosarcomaECACC Cellosaurus
HT-29HumanColon epitheliumAdenocarcinomaECACC Cellosaurus
J558LMouseMyelomaB lymphocyte cellECACC Cellosaurus
JurkatHumanWhite blood cellsT cell leukemiaECACC Cellosaurus
JYHumanLymphoblastoidEBV-transformed B cellECACC Cellosaurus
K562HumanLymphoblastoidCML blast crisisECACC Cellosaurus
KBM-7HumanLymphoblastoidCML blast crisisCellosaurus
KCL-22HumanLymphoblastoidCMLDSMZ Cellosaurus
KG1HumanLymphoblastoidAMLECACC Cellosaurus
Ku812HumanLymphoblastoidErythroleukemiaECACC Cellosaurus
KYO-1Kyoto-1HumanLymphoblastoidCMLDSMZ Cellosaurus
L1210MouseLymphocytic leukemiaAscitic fluidECACC Cellosaurus
L243MouseHybridomaSecretes L243 mAb (against HLA-DR)ATCC Cellosaurus
LNCaPLymph Node Cancer of the ProstateHumanProstatic adenocarcinomaEpithelialECACC Cellosaurus
MA-104Microbiological Associates-104African Green MonkeyKidneyEpithelialCellosaurus
MA2.1MouseHybridomaSecretes MA2.1 mAb (against HLA-A2 and HLA-B17)ATCC Cellosaurus
Ma-Mel 1, 2, 3....48HumanSkinA range of melanoma cell linesECACC Cellosaurus
MC-38Mouse Colon-38MouseColonAdenocarcinomaCellosaurus
MCF-7Michigan Cancer Foundation-7HumanBreastInvasive breast ductal carcinoma ER+, PR+ECACC Cellosaurus
MCF-10AMichigan Cancer Foundation-10AHumanBreast epitheliumATCC Cellosaurus
MDA-MB-157M.D. Anderson - Metastatic Breast-157HumanPleural effusion metastasisBreast carcinomaECACC Cellosaurus
MDA-MB-231M.D. Anderson - Metastatic Breast-231HumanPleural effusion metastasisBreast carcinomaECACC Cellosaurus
MDA-MB-361M.D. Anderson - Metastatic Breast-361HumanMelanoma (contaminated by M14)ECACC Cellosaurus
MDA-MB-468M.D. Anderson - Metastatic Breast-468HumanPleural effusion metastasisBreast carcinomaATCC Cellosaurus
MDCK IIMadin Darby Canine Kidney IIDogKidneyEpitheliumECACC Cellosaurus
MG63HumanBoneOsteosarcomaECACC Cellosaurus
MIA PaCa-2HumanProstatePancreatic CarcinomaATCC Cellosaurus
MOR/0.2RHumanLungLung carcinomaECACC Cellosaurus
Mono-Mac-6HumanWhite blood cellsMyeloid metaplasic AMLDSMZ Cellosaurus
MRC-5Medical Research Council cell strain 5HumanLung (fetal)FibroblastECACC Cellosaurus
MTD-1AMouseEpitheliumCellosaurus
MyEndMyocardial EndothelialMouseEndotheliumCellosaurus
NCI-H69HumanLungLung carcinomaECACC Cellosaurus
NCI-H69/CPRHumanLungLung carcinomaECACC Cellosaurus
NCI-H69/LX10HumanLungLung carcinomaECACC Cellosaurus
NCI-H69/LX20HumanLungLung carcinomaECACC Cellosaurus
NCI-H69/LX4HumanLungLung carcinomaECACC Cellosaurus
Neuro-2aMouseNerve/neuroblastomaNeuronal stem cellsECACC Cellosaurus
NIH-3T3NIH, 3-day transfer, inoculum 3 x 105 cellsMouseEmbryoFibroblastECACC Cellosaurus
NALM-1HumanPeripheral bloodBlast-crisis CMLATCC Cellosaurus
NK-92HumanLeukemia/lymphomaATCC Cellosaurus
NTERA-2HumanLung metastasisEmbryonal carcinomaECACC Cellosaurus
NW-145HumanSkinMelanomaESTDAB Archived 2011-11-16 at the Wayback Machine Cellosaurus
OKOpossum KidneyVirginia opossum - Didelphis virginianaKidneyECACC Cellosaurus
OPCN / OPCT cell linesHumanProstateRange of prostate tumour linesCellosaurus
P3X63Ag8MouseMyelomaECACC Cellosaurus
PANC-1HumanDuctEpithelioid CarcinomaATCC Cellosaurus
PC12RatAdrenal medullaPheochromocytomaECACC Cellosaurus
PC-3Prostate Cancer-3HumanBone metastasisProstate carcinomaECACC Cellosaurus
PeerHumanT cell leukemiaDSMZ Cellosaurus
PNT1AHumanProstateSV40-transformed tumour lineECACC Cellosaurus
PNT2HumanProstateSV40-transformed tumour lineECACC Cellosaurus
Pt K2The second cell line derived from Potorous tridactylisLong-nosed potoroo - Potorous tridactylusKidneyEpithelialECACC Cellosaurus
RajiHumanB lymphomaLymphoblast-likeECACC Cellosaurus
RBL-1Rat Basophilic Leukemia-1RatLeukemiaBasophil cellECACC Cellosaurus
RenCaRenal CarcinomaMouseKidneyRenal carcinomaATCC Cellosaurus
RIN-5FMousePancreasECACC Cellosaurus
RMA-SMouseT cell tumourCellosaurus
S2Schneider 2Insect - Drosophila melanogasterLate stage (20–24 hours old) embryosATCC Cellosaurus
SaOS-2Sarcoma OSteogenic-2HumanBoneOsteosarcomaECACC Cellosaurus
Sf21Spodoptera frugiperda 21Insect (moth) - Spodoptera frugiperdaOvaryECACC Cellosaurus
Sf9Spodoptera frugiperda 9Insect (moth) - Spodoptera frugiperdaOvaryECACC Cellosaurus
SH-SY5YHumanBone marrow metastasisNeuroblastomaECACC Cellosaurus
SiHaHumanCervix epitheliumCervical carcinomaATCC Cellosaurus
SK-BR-3Sloan-Kettering Breast cancer 3HumanBreastBreast carcinomaDSMZ Cellosaurus
SK-OV-3Sloan-Kettering Ovarian cancer 3HumanOvaryOvarian carcinomaECACC Cellosaurus
SK-N-SHHumanBrainEpithelialATCC Cellosaurus
T2HumanT cell leukemia/B cell line hybridomaATCC Cellosaurus
T-47DHumanBreastBreast ductal carcinomaECACC Cellosaurus
T84HumanLung metastasisColorectal carcinomaECACC Cellosaurus
T98GHumanGlioblastoma-astrocytomaEpitheliumECACC Cellosaurus
THP-1HumanMonocyteAcute monocytic leukemiaECACC Cellosaurus
U2OSHumanOsteosarcomaEpithelialECACC Cellosaurus
U373HumanGlioblastoma-astrocytomaEpitheliumECACC Cellosaurus
U87HumanGlioblastoma-astrocytomaEpithelial-likeECACC Cellosaurus
U937HumanLeukemic monocytic lymphomaECACC Cellosaurus
VCaPVertebral Cancer of the ProstateHumanVertebra metastasisProstate carcinomaECACC Cellosaurus
VeroFrom Esperanto: verda (green, for green monkey) reno (kidney)African green monkey - Chlorocebus sabaeusKidney epitheliumECACC Cellosaurus
VG-1HumanPrimary effusion lymphomaCellosaurus
WM39HumanSkinMelanomaESTDAB Cellosaurus
WT-49HumanLymphoblastoidECACC Cellosaurus
YAC-1MouseLymphomaECACC Cellosaurus
YARHumanLymphoblastoidEBV-transformed B cellHuman Immunology[93] ECACC Cellosaurus

See also
  • Biological immortality
  • Cell culture assays
  • Electric cell-substrate impedance sensing
  • List of contaminated cell lines
  • List of NCI-60 Cell Lines
  • List of LL-100 panel Cell Lines
  • List of breast cancer cell lines
  • Microphysiometry

References and notes
  1. Carrel A, Burrows MT (March 1911). "Cultivation of Tissues in Vitro and ITS Technique". The Journal of Experimental Medicine. 13 (3): 387–396. doi:10.1084/jem.13.3.387. PMC 2125263. PMID 19867420.
  2. Taylor MW (2014). "A History of Cell Culture". Viruses and Man: A History of Interactions. Cham: Springer International Publishing. pp. 41–52. doi:10.1007/978-3-319-07758-1_3. ISBN 978-3-319-07757-4.
  3. Harris AR, Peter L, Bellis J, Baum B, Kabla AJ, Charras GT (October 2012). "Characterizing the mechanics of cultured cell monolayers". Proceedings of the National Academy of Sciences of the United States of America. 109 (41): 16449–16454. Bibcode:2012PNAS..10916449H. doi:10.1073/pnas.1213301109. PMC 3478631. PMID 22991459.
  4. "Some landmarks in the development of tissue and cell culture". Retrieved 2006-04-19.
  5. "Cell Culture". Retrieved 2006-04-19.
  6. "Whonamedit - Ringer's solution". whonamedit.com. Retrieved 2014-06-09.
  7. Steinhardt E, Israeli C, Lambert RA (1913). "Studies on the Cultivation of the Virus of Vaccinia". The Journal of Infectious Diseases. 13 (2): 294–300. doi:10.1093/infdis/13.2.294. ISSN 0022-1899. JSTOR 30073371.
  8. Atala A (2009). "Growing new organs". TEDMED. Retrieved 2021-08-23.
  9. "Animals and alternatives in testing". Archived from the original on 2006-02-25. Retrieved 2006-04-19.
  10. Fentem JH (February 2006). "Working together to respond to the challenges of EU policy to replace animal testing". Alternatives to Laboratory Animals. 34 (1): 11–18. doi:10.1177/026119290603400116. PMID 16522146. S2CID 10339716.
  11. Schiff JA (February 2002). "An unsung hero of medical research". Yale Alumni Magazine. Archived from the original on 2012-11-14. Retrieved 2006-04-19.
  12. Bonner J (June 1936). "Plant Tissue Cultures from a Hormone Point of View". Proceedings of the National Academy of Sciences of the United States of America. 22 (6): 426–430. Bibcode:1936PNAS...22..426B. doi:10.1073/pnas.22.6.426. JSTOR 86579. PMC 1076796. PMID 16588100.
  13. Haberlandt, G. (1902) Kulturversuche mit isolierten Pflanzenzellen. Sitzungsber. Akad. Wiss. Wien. Math.-Naturwiss. Kl., Abt. J. 111, 69–92.
  14. Noé AC (October 1934). "Gottlieb Haberlandt". Plant Physiology. 9 (4): 850–855. doi:10.1104/pp.9.4.850. PMC 439112. PMID 16652925.
  15. Plant Tissue Culture. 100 years since Gottlieb Haberlandt. Laimer, Margit; Rücker, Waltraud (Eds.) 2003. Springer ISBN 978-3-211-83839-6
  16. Martin BM (2013-12-01). Tissue Culture Techniques: An Introduction. Springer Science & Business Media. pp. 29–30. ISBN 978-1-4612-0247-9.
  17. Simon EM (1988). "Phase I Final Report: Fibrous Substrates for Cell Culture (R3RR03544A)". ResearchGate. Retrieved 2017-05-22.
  18. Urry, L. A., Campbell, N. A., Cain, M. L., Reece, J. B., Wasserman, S. (2007). Biology. United Kingdom: Benjamin-Cummings Publishing Company. p. 860
  19. Voigt N, Pearman CM, Dobrev D, Dibb KM (September 2015). "Methods for isolating atrial cells from large mammals and humans". Journal of Molecular and Cellular Cardiology. 86: 187–198. doi:10.1016/j.yjmcc.2015.07.006. PMID 26186893.
  20. Louch WE, Sheehan KA, Wolska BM (September 2011). "Methods in cardiomyocyte isolation, culture, and gene transfer". Journal of Molecular and Cellular Cardiology. 51 (3): 288–298. doi:10.1016/j.yjmcc.2011.06.012. PMC 3164875. PMID 21723873.
  21. Hemeda, H., Giebel, B., Wagner, W. (16Feb2014) Evaluation of human platelet lysate versus fetal bovine serum for culture of mesenchymal stromal cells Cytotherapy p170-180 issue 2 doi.10.1016
  22. "Post - Blog | Boval BioSolutions, LLC". bovalco.com. Retrieved 2014-12-02.
  23. "LipiMAX purified lipoprotein solution from bovine serum". Selborne Biological Services. 2006. Archived from the original on 2012-07-19. Retrieved 2010-02-02.
  24. Portela VM, Zamberlam G, Price CA (April 2010). "Cell plating density alters the ratio of estrogenic to progestagenic enzyme gene expression in cultured granulosa cells". Fertility and Sterility. 93 (6): 2050–2055. doi:10.1016/j.fertnstert.2009.01.151. PMID 19324349.
  25. Jaccard N, Macown RJ, Super A, Griffin LD, Veraitch FS, Szita N (October 2014). "Automated and online characterization of adherent cell culture growth in a microfabricated bioreactor". Journal of Laboratory Automation. 19 (5): 437–443. doi:10.1177/2211068214529288. PMC 4230958. PMID 24692228.
  26. Humpel C (October 2015). "Organotypic brain slice cultures: A review". Neuroscience. 305: 86–98. doi:10.1016/j.neuroscience.2015.07.086. PMC 4699268. PMID 26254240.
  27. Neimark J (February 2015). "Line of attack". Science. 347 (6225): 938–940. Bibcode:2015Sci...347..938N. doi:10.1126/science.347.6225.938. PMID 25722392.
  28. Drexler HG, Dirks WG, MacLeod RA (October 1999). "False human hematopoietic cell lines: cross-contaminations and misinterpretations". Leukemia. 13 (10): 1601–1607. doi:10.1038/sj.leu.2401510. PMID 10516762.
  29. Drexler HG, MacLeod RA, Dirks WG (December 2001). "Cross-contamination: HS-Sultan is not a myeloma but a Burkitt lymphoma cell line". Blood. 98 (12): 3495–3496. doi:10.1182/blood.V98.12.3495. PMID 11732505.
  30. Cabrera CM, Cobo F, Nieto A, Cortés JL, Montes RM, Catalina P, Concha A (June 2006). "Identity tests: determination of cell line cross-contamination". Cytotechnology. 51 (2): 45–50. doi:10.1007/s10616-006-9013-8. PMC 3449683. PMID 19002894.
  31. Chatterjee R (February 2007). "Cell biology. Cases of mistaken identity". Science. 315 (5814): 928–931. doi:10.1126/science.315.5814.928. PMID 17303729. S2CID 13255156.
  32. Liscovitch M, Ravid D (January 2007). "A case study in misidentification of cancer cell lines: MCF-7/AdrR cells (re-designated NCI/ADR-RES) are derived from OVCAR-8 human ovarian carcinoma cells". Cancer Letters. 245 (1–2): 350–352. doi:10.1016/j.canlet.2006.01.013. PMID 16504380.
  33. MacLeod RA, Dirks WG, Matsuo Y, Kaufmann M, Milch H, Drexler HG (November 1999). "Widespread intraspecies cross-contamination of human tumor cell lines arising at source". International Journal of Cancer. 83 (4): 555–563. doi:10.1002/(SICI)1097-0215(19991112)83:4<555::AID-IJC19>3.0.CO;2-2. PMID 10508494.
  34. Masters JR (April 2002). "HeLa cells 50 years on: the good, the bad and the ugly". Nature Reviews. Cancer. 2 (4): 315–319. doi:10.1038/nrc775. PMID 12001993. S2CID 991019.
  35. Dunham JH, Guthmiller P (2008). "Doing good science: Authenticating cell line identity" (PDF). Cell Notes. 22: 15–17. Archived from the original (PDF) on 2008-10-28. Retrieved 2008-10-28.
  36. Brendan P. Lucey, Walter A. Nelson-Rees, Grover M. Hutchins; Henrietta Lacks, HeLa Cells, and Cell Culture Contamination. Arch Pathol Lab Med 1 September 2009; 133 (9): 1463–1467. doi: https://doi.org/10.5858/133.9.1463
  37. Nguyen HT, Geens M, Spits C (2012). "Genetic and epigenetic instability in human pluripotent stem cells". Human Reproduction Update. 19 (2): 187–205. doi:10.1093/humupd/dms048. PMID 23223511.
  38. Lagziel S, Gottlieb E, Shlomi T (December 2020). "Mind your media". Nature Metabolism. 2 (12): 1369–1372. doi:10.1038/s42255-020-00299-y. PMID 33046912. S2CID 222319735.
  39. Lagziel S, Lee WD, Shlomi T (April 2019). "Inferring cancer dependencies on metabolic genes from large-scale genetic screens". BMC Biology. 17 (1): 37. doi:10.1186/s12915-019-0654-4. PMC 6489231. PMID 31039782.
  40. Vande Voorde J, Ackermann T, Pfetzer N, Sumpton D, Mackay G, Kalna G, et al. (January 2019). "Improving the metabolic fidelity of cancer models with a physiological cell culture medium". Science Advances. 5 (1): eaau7314. Bibcode:2019SciA....5.7314V. doi:10.1126/sciadv.aau7314. PMC 6314821. PMID 30613774.
  41. Cantor JR, Abu-Remaileh M, Kanarek N, Freinkman E, Gao X, Louissaint A, et al. (April 2017). "Physiologic Medium Rewires Cellular Metabolism and Reveals Uric Acid as an Endogenous Inhibitor of UMP Synthase". Cell. 169 (2): 258–272.e17. doi:10.1016/j.cell.2017.03.023. PMC 5421364. PMID 28388410.
  42. "Moore v. Regents of University of California (1990) 51 C3d 120". Online.ceb.com. Retrieved 2012-01-27.
  43. Hayflick L (September 1998). "A brief history of the mortality and immortality of cultured cells". The Keio Journal of Medicine. 3. 47 (3): 174–182. doi:10.2302/kjm.47.174. PMID 9785764.
  44. "Worthington tissue guide". Retrieved 2013-04-30.
  45. Qian L, Saltzman WM (2004). "Improving the expansion and neuronal differentiation of mesenchymal stem cells through culture surface modification". Biomaterials. 25 (7–8): 1331–1337. doi:10.1016/j.biomaterials.2003.08.013. PMID 14643607.
  46. Maguire G (May 2016). "Therapeutics from Adult Stem Cells and the Hype Curve". ACS Medicinal Chemistry Letters. 7 (5): 441–443. doi:10.1021/acsmedchemlett.6b00125. PMC 4867479. PMID 27190588.
  47. Prieto D, Aparicio G, Sotelo-Silveira JR (November 2017). "Cell migration analysis: A low-cost laboratory experiment for cell and developmental biology courses using keratocytes from fish scales". Biochemistry and Molecular Biology Education. 45 (6): 475–482. doi:10.1002/bmb.21071. PMID 28627731.
  48. Discher DE, Janmey P, Wang YL (November 2005). "Tissue cells feel and respond to the stiffness of their substrate". Science. 310 (5751): 1139–1143. Bibcode:2005Sci...310.1139D. CiteSeerX 10.1.1.318.690. doi:10.1126/science.1116995. PMID 16293750. S2CID 9036803.
  49. Gilbert PM, Havenstrite KL, Magnusson KE, Sacco A, Leonardi NA, Kraft P, et al. (August 2010). "Substrate elasticity regulates skeletal muscle stem cell self-renewal in culture". Science. 329 (5995): 1078–1081. Bibcode:2010Sci...329.1078G. doi:10.1126/science.1191035. PMC 2929271. PMID 20647425.
  50. Chowdhury F, Li Y, Poh YC, Yokohama-Tamaki T, Wang N, Tanaka TS (December 2010). Zhou Z (ed.). "Soft substrates promote homogeneous self-renewal of embryonic stem cells via downregulating cell-matrix tractions". PLOS ONE. 5 (12): e15655. Bibcode:2010PLoSO...515655C. doi:10.1371/journal.pone.0015655. PMC 3001487. PMID 21179449.
  51. Engler AJ, Sen S, Sweeney HL, Discher DE (August 2006). "Matrix elasticity directs stem cell lineage specification". Cell. 126 (4): 677–689. doi:10.1016/j.cell.2006.06.044. PMID 16923388.
  52. Paszek MJ, Zahir N, Johnson KR, Lakins JN, Rozenberg GI, Gefen A, et al. (September 2005). "Tensional homeostasis and the malignant phenotype". Cancer Cell. 8 (3): 241–254. doi:10.1016/j.ccr.2005.08.010. PMID 16169468.
  53. Levental KR, Yu H, Kass L, Lakins JN, Egeblad M, Erler JT, et al. (November 2009). "Matrix crosslinking forces tumor progression by enhancing integrin signaling". Cell. 139 (5): 891–906. doi:10.1016/j.cell.2009.10.027. PMC 2788004. PMID 19931152.
  54. Tilghman RW, Cowan CR, Mih JD, Koryakina Y, Gioeli D, Slack-Davis JK, et al. (September 2010). Hotchin NA (ed.). "Matrix rigidity regulates cancer cell growth and cellular phenotype". PLOS ONE. 5 (9): e12905. Bibcode:2010PLoSO...512905T. doi:10.1371/journal.pone.0012905. PMC 2944843. PMID 20886123.
  55. Liu F, Mih JD, Shea BS, Kho AT, Sharif AS, Tager AM, Tschumperlin DJ (August 2010). "Feedback amplification of fibrosis through matrix stiffening and COX-2 suppression". The Journal of Cell Biology. 190 (4): 693–706. doi:10.1083/jcb.201004082. PMC 2928007. PMID 20733059.
  56. Wipff PJ, Rifkin DB, Meister JJ, Hinz B (December 2007). "Myofibroblast contraction activates latent TGF-beta1 from the extracellular matrix". The Journal of Cell Biology. 179 (6): 1311–1323. doi:10.1083/jcb.200704042. PMC 2140013. PMID 18086923.
  57. Georges PC, Hui JJ, Gombos Z, McCormick ME, Wang AY, Uemura M, et al. (December 2007). "Increased stiffness of the rat liver precedes matrix deposition: implications for fibrosis". American Journal of Physiology. Gastrointestinal and Liver Physiology. 293 (6): G1147–G1154. doi:10.1152/ajpgi.00032.2007. PMID 17932231. S2CID 201357.
  58. Li L, Sharma N, Chippada U, Jiang X, Schloss R, Yarmush ML, Langrana NA (May 2008). "Functional modulation of ES-derived hepatocyte lineage cells via substrate compliance alteration". Annals of Biomedical Engineering. 36 (5): 865–876. doi:10.1007/s10439-008-9458-3. PMID 18266108. S2CID 21773886.
  59. Semler EJ, Lancin PA, Dasgupta A, Moghe PV (February 2005). "Engineering hepatocellular morphogenesis and function via ligand-presenting hydrogels with graded mechanical compliance". Biotechnology and Bioengineering. 89 (3): 296–307. doi:10.1002/bit.20328. PMID 15744840.
  60. Friedland JC, Lee MH, Boettiger D (January 2009). "Mechanically activated integrin switch controls alpha5beta1 function". Science. 323 (5914): 642–644. Bibcode:2009Sci...323..642F. doi:10.1126/science.1168441. PMID 19179533. S2CID 206517419.
  61. Chan CE, Odde DJ (December 2008). "Traction dynamics of filopodia on compliant substrates". Science. 322 (5908): 1687–1691. Bibcode:2008Sci...322.1687C. doi:10.1126/science.1163595. PMID 19074349. S2CID 28568350.
  62. Dupont S, Morsut L, Aragona M, Enzo E, Giulitti S, Cordenonsi M, et al. (June 2011). "Role of YAP/TAZ in mechanotransduction". Nature. 474 (7350): 179–183. doi:10.1038/nature10137. hdl:11380/673649. PMID 21654799. S2CID 205225137.
  63. "drug discovery@nature.com". Nature.com. Retrieved 2013-03-26.
  64. Duell BL, Cripps AW, Schembri MA, Ulett GC (2011). "Epithelial cell coculture models for studying infectious diseases: benefits and limitations". Journal of Biomedicine & Biotechnology. 2011: 852419. doi:10.1155/2011/852419. PMC 3189631. PMID 22007147.
  65. Barrila J, Radtke AL, Crabbé A, Sarker SF, Herbst-Kralovetz MM, Ott CM, Nickerson CA (November 2010). "Organotypic 3D cell culture models: using the rotating wall vessel to study host-pathogen interactions". Nature Reviews. Microbiology. 8 (11): 791–801. doi:10.1038/nrmicro2423. PMID 20948552. S2CID 6925183.
  66. Mapanao AK, Voliani V (June 2020). "Three-dimensional tumor models: Promoting breakthroughs in nanotheranostics translational research". Applied Materials Today. 19: 100552. doi:10.1016/j.apmt.2019.100552. S2CID 213634060.
  67. Cassano D, Santi M, D'Autilia F, Mapanao AK, Luin S, Voliani V (2019). "Photothermal effect by NIR-responsive excretable ultrasmall-in-nano architectures". Materials Horizons. 6 (3): 531–537. doi:10.1039/C9MH00096H. ISSN 2051-6347.
  68. Edmondson R, Broglie JJ, Adcock AF, Yang L (May 2014). "Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors". Assay and Drug Development Technologies. 12 (4): 207–218. doi:10.1089/adt.2014.573. PMC 4026212. PMID 24831787.
  69. Bhattacharya M, Malinen MM, Lauren P, Lou YR, Kuisma SW, Kanninen L, et al. (December 2012). "Nanofibrillar cellulose hydrogel promotes three-dimensional liver cell culture". Journal of Controlled Release. 164 (3): 291–298. doi:10.1016/j.jconrel.2012.06.039. PMID 22776290.
  70. DeRosa MC, Monreal C, Schnitzer M, Walsh R, Sultan Y (February 2010). "Nanotechnology in fertilizers". Nature Nanotechnology. 5 (2): 91. Bibcode:2010NatNa...5...91D. doi:10.1038/nnano.2010.2. PMID 20130583.
  71. Hsiao AY, Tung YC, Qu X, Patel LR, Pienta KJ, Takayama S (May 2012). "384 hanging drop arrays give excellent Z-factors and allow versatile formation of co-culture spheroids". Biotechnology and Bioengineering. 109 (5): 1293–1304. doi:10.1002/bit.24399. PMC 3306496. PMID 22161651.
  72. Mapanao AK, Santi M, Faraci P, Cappello V, Cassano D, Voliani V (September 2018). "Endogenously Triggerable Ultrasmall-in-Nano Architectures: Targeting Assessment on 3D Pancreatic Carcinoma Spheroids". ACS Omega. 3 (9): 11796–11801. doi:10.1021/acsomega.8b01719. PMC 6173554. PMID 30320273.
  73. Ghosh S, Börsch A, Ghosh S, Zavolan M (April 2021). "The transcriptional landscape of a hepatoma cell line grown on scaffolds of extracellular matrix proteins". BMC Genomics. 22 (1): 238. doi:10.1186/s12864-021-07532-2. PMC 8025518. PMID 33823809.
  74. Fontoura JC, Viezzer C, Dos Santos FG, Ligabue RA, Weinlich R, Puga RD, et al. (February 2020). "Comparison of 2D and 3D cell culture models for cell growth, gene expression and drug resistance". Materials Science & Engineering. C, Materials for Biological Applications. 107: 110264. doi:10.1016/j.msec.2019.110264. PMID 31761183. S2CID 208277016.
  75. Habanjar O, Diab-Assaf M, Caldefie-Chezet F, Delort L (November 2021). "3D Cell Culture Systems: Tumor Application, Advantages, and Disadvantages". International Journal of Molecular Sciences. 22 (22): 12200. doi:10.3390/ijms222212200. PMC 8618305. PMID 34830082.
  76. Tibbitt MW, Anseth KS (July 2009). "Hydrogels as extracellular matrix mimics for 3D cell culture". Biotechnology and Bioengineering. 103 (4): 655–663. doi:10.1002/bit.22361. PMC 2997742. PMID 19472329.
  77. "Quickie Bird Flu Vaccine Created". Wired. Wired.com. Reuters. 2006-01-26. Retrieved 2010-01-31.
  78. Gao W, Soloff AC, Lu X, Montecalvo A, Nguyen DC, Matsuoka Y, et al. (February 2006). "Protection of mice and poultry from lethal H5N1 avian influenza virus through adenovirus-based immunization". Journal of Virology. 80 (4): 1959–1964. doi:10.1128/JVI.80.4.1959-1964.2006. PMC 1367171. PMID 16439551.
  79. "NIAID Taps Chiron to Develop Vaccine Against H9N2 Avian Influenza". National Institute of Allergy and Infectious Diseases (NIAID). 2004-08-17. Retrieved 2010-01-31.
  80. Miki, Yasuhiro; Ono, Katsuhiko; Hata, Shuko; Suzuki, Takashi; Kumamoto, Hiroyuki; Sasano, Hironobu (September 2012). "The advantages of co-culture over mono cell culture in simulating in vivo environment". The Journal of Steroid Biochemistry and Molecular Biology. 131 (3–5): 68–75. doi:10.1016/j.jsbmb.2011.12.004. ISSN 0960-0760.
  81. Paschos, Nikolaos K.; Brown, Wendy E.; Eswaramoorthy, Rajalakshmanan; Hu, Jerry C.; Athanasiou, Kyriacos A. (2014-02-03). "Advances in tissue engineering through stem cell-based co-culture". Journal of Tissue Engineering and Regenerative Medicine. 9 (5): 488–503. doi:10.1002/term.1870. ISSN 1932-6254.
  82. Dittrich, Petra S.; Manz, Andreas (March 2006). "Lab-on-a-chip: microfluidics in drug discovery". Nature Reviews Drug Discovery. 5 (3): 210–218. doi:10.1038/nrd1985. ISSN 1474-1784.
  83. A. Terrell, John; G. Jones, Curtis; Monia Kabandana, Giraso Keza; Chen, Chengpeng (2020). "From cells-on-a-chip to organs-on-a-chip: scaffolding materials for 3D cell culture in microfluidics". Journal of Materials Chemistry B. 8 (31): 6667–6685. doi:10.1039/D0TB00718H. {{cite journal}}: no-break space character in |last2= at position 3 (help); no-break space character in |last3= at position 6 (help); no-break space character in |last= at position 3 (help)
  84. Wu, Qirui; Liu, Jinfeng; Wang, Xiaohong; Feng, Lingyan; Wu, Jinbo; Zhu, Xiaoli; Wen, Weijia; Gong, Xiuqing (2020-02-12). "Organ-on-a-chip: recent breakthroughs and future prospects". BioMedical Engineering OnLine. 19 (1): 9. doi:10.1186/s12938-020-0752-0. ISSN 1475-925X. PMC 7017614. PMID 32050989.
  85. Leung, Chak Ming; de Haan, Pim; Ronaldson-Bouchard, Kacey; Kim, Ge-Ah; Ko, Jihoon; Rho, Hoon Suk; Chen, Zhu; Habibovic, Pamela; Jeon, Noo Li; Takayama, Shuichi; Shuler, Michael L.; Vunjak-Novakovic, Gordana; Frey, Olivier; Verpoorte, Elisabeth; Toh, Yi-Chin (2022-05-12). "A guide to the organ-on-a-chip". Nature Reviews Methods Primers. 2 (1): 1–29. doi:10.1038/s43586-022-00118-6. ISSN 2662-8449.
  86. Ma, Chao; Peng, Yansong; Li, Hongtong; Chen, Weiqiang (February 2021). "Organ-on-a-Chip: A New Paradigm for Drug Development". Trends in Pharmacological Sciences. 42 (2): 119–133. doi:10.1016/j.tips.2020.11.009. PMC 7990030. PMID 33341248.
  87. Rapanan JL, Cooper KE, Leyva KJ, Hull EE (August 2014). "Collective cell migration of primary zebrafish keratocytes". Experimental Cell Research. 326 (1): 155–165. doi:10.1016/j.yexcr.2014.06.011. PMID 24973510.
  88. Lee J, Jacobson K (November 1997). "The composition and dynamics of cell-substratum adhesions in locomoting fish keratocytes". Journal of Cell Science. 110 (22): 2833–2844. doi:10.1242/jcs.110.22.2833. PMID 9427291.
  89. Drugmand JC, Schneider YJ, Agathos SN (2012). "Insect cells as factories for biomanufacturing". Biotechnology Advances. 30 (5): 1140–1157. doi:10.1016/j.biotechadv.2011.09.014. PMID 21983546.
  90. Hunt P, Robertson D, Weiss D, Rennick D, Lee F, Witte ON (March 1987). "A single bone marrow-derived stromal cell type supports the in vitro growth of early lymphoid and myeloid cells". Cell. 48 (6): 997–1007. doi:10.1016/0092-8674(87)90708-2. PMID 2435412. S2CID 31499611.
  91. van den Berg-Bakker CA, Hagemeijer A, Franken-Postma EM, Smit VT, Kuppen PJ, van Ravenswaay Claasen HH, et al. (February 1993). "Establishment and characterization of 7 ovarian carcinoma cell lines and one granulosa tumor cell line: growth features and cytogenetics". International Journal of Cancer. 53 (4): 613–620. doi:10.1002/ijc.2910530415. PMID 8436435. S2CID 6182244.
  92. Lee YG, Korenchuk S, Lehr J, Whitney S, Vessela R, Pienta KJ (2001). "Establishment and characterization of a new human prostatic cancer cell line: DuCaP". In Vivo. 15 (2): 157–162. PMID 11317521.
  93. Ou D, Mitchell LA, Décarie D, Tingle AJ, Nepom GT (March 1998). "Promiscuous T-cell recognition of a rubella capsid protein epitope restricted by DRB1*0403 and DRB1*0901 molecules sharing an HLA DR supertype". Human Immunology. 59 (3): 149–157. doi:10.1016/S0198-8859(98)00006-8. PMID 9548074.

Further reading
Library resources about
Cell culture
This article is issued from Wikipedia. The text is licensed under Creative Commons - Attribution - Sharealike. Additional terms may apply for the media files.