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A Serum-Free Medium for Clonal Growth and Serial Subculture of Diploid Rat Liver Epithelial Cells
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A Serum-Free Medium for Clonal Growth and Serial Subculture of Diploid Rat Liver Epithelial Cells
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A Serum-Free Medium for Clonal Growth and Serial Subculture of Diploid Rat Liver Epithelial Cells
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Journal Article
A Serum-Free Medium for Clonal Growth and Serial Subculture of Diploid Rat Liver Epithelial Cells
1983
Overview
Clonal growth and serial subculture of diploid liver epithelial cells from neonatal rats were achieved in a serum-free medium (SFM) supplemented with linoleic and oleic acid linked to fatty acid-free bovine serum albumin (fafBSA), epidermal growth factor (EGF), transferrin, insulin, selenous acid, and fetuin. Because it is not known whether factors added to defined media facilitate attachment, support proliferation, or both, a serum-free \"attachment medium\" was first devised in which cells would attach to the substratum without loss of viability. Then a growth medium that would support cell proliferation was developed. Fetuin enhanced the degree of attachment, and the lipid supplements and EGF induced a marked proliferative response. Serum-free medium supported the formation of colonies equivalent in size, number, and morphology to those obtained in serum-supplemented medium. Cells plated at a higher inoculum density and subcultured regularly for up to 25 wk underwent two to three doublings per week and acquired a flattened epithelial cell morphology. Early passages of rat liver epithelial cells, cultured in SFM may be useful in studies of the regulation of cell proliferation and differentiation.
Publisher
Tissue Culture Association, Inc,Tissue Culture Association
Subject
alpha-Fetoproteins - pharmacology
/ Animals
/ Biological and medical sciences
/ Cell Adhesion - drug effects
/ Cell cultures. Hybridization. Fusion
/ Cell Division - drug effects
/ Diploidy
/ Epidermal Growth Factor - pharmacology
/ Establishment of new cell lines, improvement of cultural methods, mass cultures
/ Fundamental and applied biological sciences. Psychology
/ Insulin
/ Liver
/ Methods. Procedures. Technologies
/ Molecular and cellular biology
/ Rats
