Summary
Mouse fibroblasts were cultured by three different procedures: (a) changing the 0.2 ml/cm2 of growth medium every 2nd d and seeding 1×105 cells/cm2 after confluency; (b) changing the 0.4 ml/cm2 of growth medium only at subculture performed at confluency by a 1∶2 split and keeping the bottles incubated on a rocking platform; (c) the same as Method b but keeping the bottles stationary throughout culture. By Method a no lines were immortalized over 36 experiments whereas Method b gave 1/4 immortalized lines and Method c gave 10∶12 immortalized lines. Cells always went into crisis at the 9th to 11th doubling. Immortalized lines had a tetraploid DNA content.
Similar content being viewed by others
References
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.
Macieira-Coelho, A.; Diatloff, C.; Malaise, E. Effect of low dose rate irradiation on the division potential of cells in vitro. II. Mouse lung fibroblasts. Exp. Cell Res. 100: 228–232; 1976.
Aaronson, S. A.; Todaro, G. J. Development of 3T3-like lines from BALB/c mouse embryo cultures: Transformation susceptibility to SV40. J. Cell. Physiol. 72: 141–148; 1968.
Curatolo, L.; Balconi, G.; Borgia, R.; Morasca, L.; Donati, M. B. Fibrin clot retractile activity of mouse fibroblasts during growth and aging. In Vitro 16: 731–737; 1980.
Fogh, J.; Fogh, H. A method for direct demonstration of pleuropneumonia-like organisms in cultured cells. Proc. Soc. Exp Biol. Med. 117: 899–901; 1964.
Lau, C. C.; Pardee, A. B. Mechanism by which caffeine potentiates lethality of nitrogen mustard. Proc. Natl. Acad. Sci. USA 79: 2942–2946; 1982.
Baisch, H.; Göhde, W.; Linden, W. A. Analysis of PCP-data to determine the fraction of cells in the various phases of cell cycle. Radiat. Environ. Biophys. 12: 31–39; 1975.
Thrash, C. R.; Cunningham, D. D. Growth limitation of 3T3 mouse fibroblasts by available growth surface area and medium components. J. Cell. Physiol. 86: 301–310; 1974.
Balin, A. K.; Goodman, D. B. P.; Rasmussen, H.; Cristofalo, V. J. The effect of oxygen tension on the growth and metabolism of WI-38 cells. J. Cell. Physiol. 89: 235–250; 1976.
Goldblatt, H.; Friedman, L.; Cechner, R. L. On the malignant transformation of cells during prolonged culture under hypoxic conditions in vitro. Biochem. Med. 7: 241–252; 1973.
Goldblatt, H.; Friedman, L. Prevention of malignant change in mammalian cells during prolonged culture in vitro. Proc. Natl. Acad. Sci. USA 71: 1780–1782; 1974.
Bell, E.; Marek, L. F.; Levinstone, D. S.; Merrill, C.; Sher, S.; Young, I. T.; Eden, M. Loss of division potential in vitro: Aging or differentiation? Science 202: 1158–1163; 1978.
Vaghi, M.; Curatolo, L.; Algeri, S.; Brunello, N.; Mennini, T.; Morasca, L.; Salmona, M.; Mella, L.; Locati, M.; Dolfini, E. Studies on a neuronal-like transport system for serotonin in two cell lines. Pharmacol. Res. Commun. 14: 851–859; 1982.
Curatolo, L.; Borgia, R.; Donati, M. B.; Morasca, L. Fibrin clot retractile (FCR) activity of fibroblast-like cells from normal individuals and cancer patients. Cell Biol. Int. Rep. 6: 609–618; 1982.
Azzarone, B.; Macieira-Coelho, A.; Curatolo, L.; Brouty-Boyé, D.; Vainier, O.; Donati, M. B.; Morasca, L. Fibrin clot retractile activity in normal, established and tumorigenic human epithelial cells in culture. J. Natl. Cancer Inst. 71: 879–885; 1983.
Azzarone, B.; Curatolo, L.; Carloni, G.; Donati, M. B.; Morasca, L.; Macieira-Coelho, A. Fibrin clot retraction in normal and transformed avian fibroblasts. J. Natl. Cancer Inst. 67: 89–94; 1981.
Curatolo, L.; Azzarone, B.; Fally-Crépin, C.; Morasca, L.; Macieira-Coelho, A. Comparison of fibrin clot retraction with other transformation parameters after hybridization of normal and established cell lines. Int. J. Cancer 31: 249–255; 1983.
Newbold, R. F.; Overell, R. W. Fibroblast immortality is a prerequisite for transformation by EJ c-Ha-ras oncogene. Nature 304: 648–651; 1983.
Author information
Authors and Affiliations
Additional information
This work was partially supported by CNR CT No. 82.00183.04. Judy Baggott, Felice and Vincenzo de Ceglie, Flavia Boniardi, and Antonella Bottazzi helped prepare this manuscript.
Rights and permissions
About this article
Cite this article
Curatolo, L., Erba, E. & Morasca, L. Culture conditions induce the appearance of immortalized C3H mouse cell lines. In Vitro 20, 597–601 (1984). https://doi.org/10.1007/BF02619607
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02619607