Growth of H-35 rat hepatoma cells in unsupplemented serum-free media: effect of transferrin, insulin and cell density.
Serum-free tissue culture medium consisting of a 1:1 mixture of Dulbecco's modified Eagle's medium (DMEM) and Ham's F12 medium is herein shown to support growth of Reuber H-35 cells over several days in culture. Cells were initially plated in serum containing DMEM medium for 3 h. After cell attachment, serum is removed and replaced with a serum-free 1:1 mixture of these two commercially available tissue culture media. The doubling time of cell growth in this unsupplemented serum-free medium was 46 h in lightly plated cultures over the first 5 d. The presence of transferrin (5 micrograms/ml) and insulin (3.3 nM) results in a cell doubling time of 17 h, which equaled the growth rate in medium containing 10% fetal bovine serum. In the absence of transferrin, growth rates in serum-free medium were correlated with the cell density of cultures. Conditioned medium from dense, serum-free cultures has growth-stimulating activity in recipient lightly plated cultures. This simple, serum-free culture medium will facilitate studies on the growth regulation of H-35 rat hepatoma cells.
Shapiro
L E
LE
Department of Medicine, Montefiore Medical Center, Bronx, New York.
Wagner
N
N
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D003470:Culture Media; D006133:Growth Substances; D007328:Insulin / Q000494:pharmacology*; D008114:Liver Neoplasms, Experimental / Q000473:pathology*; D051381:Rats; D014168:Transferrin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D003470:Culture Media; D006133:Growth Substances; D007328:Insulin; D014168:Transferrin