A procedure for the serum-free growth of normal human diploid fibroblasts

Methods in Cell Science - Tập 6 - Trang 123-126 - 1980
Paul D. Phillips1, Vincent J. Cristofalo1
1The Wistar Institute of Anatomy and Biology, Philadelphia, USA

Tóm tắt

We have developed a method for the growth of WI-38 cells in a serum-free medium. Basal medium MCDB-104 is supplemented with platelet-derived growth factor (30 µg/ml), epidermal growth factor (100 ng/ml), insulin (5 µg/ml), transferrin (5 µg/ml), and dexamethasone (55 ng/ml). With this medium cells will grow at a rate and to an extent similar to that produced by medium containing 10% serum. During one growth cycle the cultures can accomplish up to seven population doublings.

Tài liệu tham khảo

Phillips, P. D.; Cristofalo, V. J. Growth regulation of WI-38 cells in a serum-free medium. Exp. Cell Res. In press. Cristofalo, V. J.; Sharf, B. B. Cellular senescence and DNA synthesis. Thymidine incorporation as a measure of population age in human diploid cells. Exp. Cell Res. 76: 419–427; 1973. Cristofalo, V. J.; Charpentier, R. A standard procedure for cultivating human diploid fibroblasts to study cellular aging. J. Tissue Cult. Methods 6: 117–122; 1980. McKeehan, W. L.; McKeehan, K. A.; Hammond, S. L.; Ham, R. G. Improved medium for clonal growth of human diploid fibroblasts at low concentrations of serum protein. In Vitro 13: 399–416; 1977. Walthall, B. J.; Ham, R. G. Multiplication of human diploid fibroblasts in a synthetic medium supplemented with EGF, insulin and dexamethasone. Exp. Cell Res. In press. Bettger, W. J.; Boyce, S. T.; Walthall, B. J.; Ham, R. G. Rapid clonal growth and serial passage of human diploid fibroblasts in a lipid-enriched, synthetic medium supplemented with EGF, insulin and dexamethasone. Proc. Natl. Acad. Sci. USA. In press.