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In vitro proliferation and lifespan of human diploid fibroblasts in serum‐free BSA‐containing medium
Author(s) -
Kan Mikio,
Yamane Isao
Publication year - 1982
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041110206
Subject(s) - bovine serum albumin , fetal bovine serum , chemically defined medium , transferrin , wi 38 , growth medium , hypoxanthine , epidermal growth factor , in vitro , biochemistry , growth factor , chemistry , cell culture , biology , microbiology and biotechnology , ploidy , genetics , receptor , bacteria , gene , enzyme
We have developed two serum‐free chemically defined media (RITC 78‐6 and RITC 80‐7) that support the growth in culture of human diploid fibroblasts to the same extent as Eagle's basal medium (BME) supplemented with 10% fetal bovine serum (FBS). These two media contain modified Eagle's minimum essential medium (MEM) supplemented with nonessential amino acids, various trace metals, organic compounds and growth factors [insulin, mouse epidermal growth factor (m‐EGF), transferrin and triiodothyronine (T 3 )]. RITC 80‐7 medium differs from RITC 78‐6 in that it contains thymidine, hypoxanthine, and vitamin B 12 and supports the long‐term serial cultivation of human diploid cultures. The addition of commercial bovine serum albumin (BSA, 5 g/liter) to the medium enhances cell growth. This effect is not observed if BSA is first delipidized, but reconstitution of BSA with certain lipids restores its ability to promote growth. BSA has an inhibitory effect on cellular attachment but this is overcome when fibronectin (FN, 10 mg/liter) is added to the medium.