Modification of MCDB 110 medium to support prolonged growth and consistent high cloning efficiency of diploid human fibroblasts
- Michigan State Univ., East Lansing (USA)
In preparation for studies on the growth factor requirements of normal and transformed human fibroblasts, we have developed a serum-free medium that supports vigorous long-term serial subculture of diploid human fibroblasts and allows them to form large-sized colonies with high efficiency (40 to 60%) when plated at cloning density. This medium, which is a modification of Ham's MCDB 110 base medium with its serum replacement supplements, is relatively easy to prepare and the cost of the serum replacements is approximately the same as that of fetal bovine serum supplied at 10%. The ingredients of Supplement B of MCDB 110 medium were added in an ethanol solution, rather than in the form of liposomes, and were combined with bovine serum albumin, a lipid carrier. Gelatin and fetuin were included as attachment factors instead of polylysine. Bioassays indicated that none of the ingredients in the medium were contaminated with either epidermal growth factor or platelet-derived growth factor. In this modified serum-free medium, which the authors have designated McM + SR{sub 1}, diploid human fibroblasts grew for 21 days at the same rate as in the base medium, McM, supplemented wt 10% FBS. During the next 20 days, they underwent 15 population doublings which was 75% of the rate of cells growing in the medium containing serum.
- OSTI ID:
- 6531105
- Journal Information:
- Experimental Cell Research; (USA), Vol. 172:2; ISSN 0014-4827
- Country of Publication:
- United States
- Language:
- English
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