KBM-7 cells
KBM-7 cells are a chronic myelogenous leukemia (CML) cell line used for biomedical research. Like all cancer cell lines, it is immortal and can divide indefinitely. A unique aspect of the KBM-7 cell line is that it is near-haploid, meaning it contains only one copy for most of its chromosomes.[1] Human chromosomes are typically diploid, meaning that there are two copies of each chromosome.
Origin
KBM-7 cells were derived from a 39-year-old man with chronic myeloid leukemia in blast crisis.[1] The original cell line contained both near haploid and hyperdiploid clones. Subsequent subcloning yielded a pure near-haploid cell line.[2] Genome analysis has revealed that besides the disomic chromosome 8, a 30 megabase fragment of chromosome 15 is present in two copies.[3] Like other CML cells lines (e.g., K562) KBM-7 cells are positive for the Philadelphia chromosome harboring the BCR-ABL oncogenic fusion. KBM-7 cells have been reprogrammed to yield the HAP1 cell line which is also monosomic for chromosome 8.[4]
Cultivation and applications
KBM-7 cells grow in suspension and are maintained in Iscove's Modified Dulbecco's Medium (IMDM) supplemented with 10% fetal bovine serum. They divide approximately every 24 hours.
KBM-7 has found applications in a variety of genomic research studies; the cell line has been examined in gene silencing experiments, been reprogrammed to become a stem cell line, and served as a test model for novel drug candidates.[5]
Significance
One method of studying gene function involves "knocking out" the gene by inducing a mutation. This causes the resulting gene product to be nonfunctional, and researchers can then see how this effects the cell's function as a whole. Many gene editing procedures have very low efficiency, and often both copies of mammalian chromosomes must be knocked out in order to see a phenotypic effect. Having a near-haploid cell line such as KBM-7 greatly increases the efficiency of these studies because there is only one gene that must be knocked out. [6]
References
- Andersson, B. S., Beran, M., Pathak, S., Goodacre, A., Barlogie, B., and McCredie, K. B. (1987). "Ph-positive chronic myeloid leukemia with near-haploid conversion in vivo and establishment of a continuously growing cell line with similar cytogenetic pattern". Cancer Genet. Cytogenet. 24 (2): 335–343. doi:10.1016/0165-4608(87)90116-6. PMID 3466682.
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: CS1 maint: multiple names: authors list (link) - Kotecki M, Reddy PS, Cochran BH (1999). "Isolation and characterization of a near-haploid human cell line". Exp. Cell Res. 252 (2): 273–80. CiteSeerX 10.1.1.24.783. doi:10.1006/excr.1999.4656. PMID 10527618.
- Bürckstümmer T, Banning C, Hainzl P, Schobesberger R, Kerzendorfer C, Pauler FM, Chen D, Them N, Schischlik F, Rebsamen M, Smida M, Fece de la Cruz F, Lapao A, Liszt M, Eizinger B, Guenzl PM, Blomen VA, Konopka T, Gapp B, Parapatics K, Maier B, Stöckl J, Fischl W, Salic S, Taba Casari MR, Knapp S, Bennett KL, Bock C, Colinge J, Kralovics R, Ammerer G, Casari G, Brummelkamp TR, Superti-Furga G, Nijman SM (2013). "A reversible gene trap collection empowers haploid genetics in human cells". Nat Methods. 10 (10): 965–71. doi:10.1038/nmeth.2609. PMC 6342250. PMID 24161985.
- Carette JE, Raaben M, Wong AC, Herbert AS, Obernosterer G, Mulherkar N, Kuehne AI, Kranzusch PJ, Griffin AM, Ruthel G, Dal Cin P, Dye JM, Whelan SP, Chandran K, Brummelkamp TR (2011). "Ebola virus entry requires the cholesterol transporter Niemann-Pick C1". Nature. 477 (7364): 340–3. Bibcode:2011Natur.477..340C. doi:10.1038/nature10348. PMC 3175325. PMID 21866103.
- Essletzbichler, Patrick; et al. (December 2014). "Megabase-scale deletion using CRISPR/Cas9 to generate a fully haploid human cell line". Genome Research. 24 (12): 2059–206. doi:10.1101/gr.177220.114. PMC 4248322. PMID 25373145.
- Duncan, Lidia M.; Timms, Richard T.; Zavodszky, Eszter; Cano, Florencia; Dougan, Gordon; Randow, Felix; Lehner, Paul J. (2012-06-22). "Fluorescence-Based Phenotypic Selection Allows Forward Genetic Screens in Haploid Human Cells". PLOS ONE. 7 (6): e39651. Bibcode:2012PLoSO...739651D. doi:10.1371/journal.pone.0039651. ISSN 1932-6203. PMC 3382162. PMID 22745803.