Summary
The effect of feeder layers on cloning efficiency of transformed human cells was investigated. Embryonic human skin or lung fibroblasts; adult human skin fibroblasts; early passage cells from embryos of mouse, rat, and hamster; established mouse cell lines; 3T3 and 10T1/2 were used as feeder layers after they were lethally exposed to Co-60 gamma-rays at 3,000 rad. As test cells to study the effect of feeder layers on cloning efficiency, WI-38 CT-1 cells transformed in vitro by Co-60 gamma-rays and HGC cells cultured from a human gastric cancer were used. The effect of feeder layers on the cloning efficiency of the test cells was dependent on cell density of feeder layer cells, sources of the feeder layer cells, and kinds of test cells. An optimal density of feeder cels produced cloning efficiencies 3 to 15 times higher than in cultures without a feeder layer. Generally, high density of cells in feeder layers decreased the cloning efficiency of the test cells, presumably owing to contact inhibition of growth and depletion of essential nutrients by the feeder layer cells. Regarding the effect of the feeder layers made of human fibroblasts, there were no significant differences in population doubling levels; tissue origins of fibroblasts; or fibroblasts derived from normal individuals, patients with cancer, or with a genetically high familial incidence of cancer, hereditary adenomatosis of the colon and rectum.
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References
Puck, T. T.; Marcus, P. I.; Cieciura, S. J. Clonal growth of mammalian cells in vitro. Growth characteristics of colonies from single HeLa cells with and without a “feeder” layer. J. Exp. Med. 103: 273–283; 1956.
Rheinwald, J. G.; Green, H. Serial cultivation of strains of human epidermal keratinocytes: The formation of keratinizing colonies from single cells. Cell 6: 331–344; 1975.
Peehl, D. M.; Ham, R. G. Growth and differentiation of human keratinocytes without a feeder layer or conditioned medium. In Vitro 16: 516–525; 1980.
Marquardt, H.; Heidelberger, C. Influence of “Feeder cells” and inducers and inhibitors of microsomal mixed-function oxidases on hydrocarbon-induced malignant transformation of cells derived from C3H mouse prostate. Cancer Res. 32: 721–725; 1972.
Huberman, E.; Sachs, L. Cell-mediated mutagenesis of mammalian cells with chemical carcinogens. Int. J. Cancer 13: 326–333; 1974.
Hayflick, L. The limited in vitro lifetime of human diploid cell strains. Exp. Cell Res. 37: 614–636; 1965.
Nichols, W. W.; Murphy, D. G.; Cristofalo, V. J.; Toji, L. H.; Green, A. E.; Dwight, S. A. Characterization of a new human diploid cell strain, IMR-90. Science 196: 60–63; 1977.
Todaro, G. J.; Green, H. Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J. Cell Biol. 17: 299–313; 1963.
Reznikoff, C. A.; Brankow, D. W.; Heidelberger, C. Establishment and characterization of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. Cancer Res. 33: 3231–3238; 1973.
Matsumura, T.; Mitsui, Y.; Fujiwara, Y.; Ishii, T.; Shimada, H. Cell, tissue and organ banks in Japan with special reference to the study of premature aging. Jpn. J. Exp. Med. 50: 321–328; 1980.
Girardi, A. J.; Weinstein, D.; Moorhead, P. S. SV 40 transformation of human diploid cells — A parallel study of viral and karyologic parameters. Ann. Med. Exp. Biol. Fenn. 44: 242–254; 1966.
Namba, M.; Nishitani, K.; Kimoto, T. Carcinogenesis in tissue culture 29: Neoplastic transformation of a normal human diploid cell strain, WI-38, with Co-60 gamma-rays. Jpn. J. Exp. Med. 48: 303–311; 1978.
Namba, M.; Nishitani, K.; Kimoto, T. Characteristics of WI-38 cells (WI-38 CT-1) transformed by treatment with Co-60 gamma-rays. Gan 71: 300–307; 1980.
Nishitani, K.; Namba, M.; Kimoto, T. Lactate dehydrogenase isozyme patterns of normal human fibroblasts and their in vitro transformed counterparts by treatment with Co-60 gamma rays, SV40, or 4-nitroquinoline 1-oxide. Gan 72: 300–304; 1981.
Namba, M.; Nishitani, K.; Fukushima, F.; Kimoto, T.; Utsunomiya, J.; Hayflick, L. Neoplastic transformation of human diploid fibroblasts treated with chemical carcinogens and Co-60 gamma-rays. Gann Monogr. Cancer Res. 27: 223–231; 1981.
Akagi, T.; Kimoto, T. Human cell line (HGC-27) derived from the metastatic lymph node of gastric cancer. Acta Med. Okayama 30: 215–217; 1976.
Hayflick, L. Tissue cultures and mycoplasmas. Texas Rep. Biol. Med. 23: 285–303; 1965.
Hayflick, L.; Moorhead, P. S. The serial cultivation of human diploid cell strains. Exp. Cell Res. 25: 585–621; 1961.
Stoker, M. G. P.; Rubin, H. Density dependent inhibition of cell growth in culture. Nature 215: 171–172; 1967.
Steck, P. A.; Voss, P. G.; Wang, J. L. Growth control in culture 3T3 fibroblasts: Assays of cell proliferation and demonstration of a growth inhibitory activity. J. Cell Biol. 83: 562–575; 1979.
Peehl, D. M.; Ham, R. G. Clonal growth of human keratinocytes with small amounts of dialyzed serum. In Vitro 16: 526–540; 1980.
Green, H. Cyclic AMP in relation to proliferation of the epidermal cell: a new view. Cell 15: 801–811; 1978.
Eisinger, M.; Lee, J. S.; Hefton, J. M.; Darzynkiewicz, Z.; Chiao, J. W.; de Harven, E. Human epidermal cell cultures: Growth and differentiation in the absence of dermal components or medium supplements. Proc. Natl. Acad. Sci. USA 76: 5340–5344; 1979.
Hawley-Nelson, P.; Sullivan, J. E.; Kung, M.; Hennings, H.; Yuspa, S. H. Optimized conditions for the growth of human epidermal cells in culture. J. Invest. Dermatol. 75: 176–182; 1980.
Lechner, J.; Babcock, M. S.; Marnell, M.; Narayan, S.; Kaighn, M. E. Normal human prostate epithelial cell cultures. Harris, C. C.; Trump, B. F.; Stoner, G. D. eds. Methods in cell biology. Vol. 21. New York: Academic Press; 1980: 195–225.
Stoner, G. D.; Katoh, Y.; Foidart, J. M.; Trump, B. F.; Steinert, P. M.; Harris, C. C. Cultured human bronchial epithelial cells: Blood group antigens, keratin, collagens, and fibronectin. In Vitro 17: 577–587; 1981.
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This study was supported by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science and Culture, Japan.
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Namba, M., Fukushima, F. & Kimoto, T. Effects of feeder layers made of human, mouse, hamster, and rat cells on the cloning efficiency of transformed human cells. In Vitro Cell.Dev.Biol.-Plant 18, 469–475 (1982). https://doi.org/10.1007/BF02796475
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DOI: https://doi.org/10.1007/BF02796475