Substrate-dependent differences in growth and biological properties of fibroblasts and epithelial cells grown in microcarrier culture.
Normal diploid human fibroblasts and first passage monkey kidney epithelial cells were examined for growth and metabolic activity on microcarriers made from glass and on microcarriers made from DEAE-dextran. The cells grew to a higher density (cells cm2 of surface area) on the glass microcarriers made from glass and on microcarriers made from DEAE-dextran. The cells grew to a higher density (cells/cm2 of surface area) on the glass microcarriers than they did on the DEAE-dextran microcarriers and morphological differences were observed between the cells growing on the two substrates. On the DEAE-dextran microcarriers, the cells were much more resistant to protease-mediated detachment than were the cells on the glass microcarriers. In these respects, the cells grown on the glass microcarriers were similar to cells grown in conventional monolayer culture. Interestingly, the cells grown on the DEAE-dextran microcarriers expressed higher levels of proteolytic enzyme activity than the cells grown on the glass microcarriers. Substrate-dependent differences in prostaglandin production also occurred--both in unstimulated cells and in cells stimulated with 12-0-tetradecanoyl phorbol acetate. The unstimulated cells on the glass microcarriers produced slightly higher levels of three different prostaglandins than did the cells on the DEAE-dextran microcarriers. However, after stimulation the levels were much higher in the DEAE-dextran microcarrier cultures than in the glass microcarrier cultures. In contrast to these results, there was no significant, substrate-dependent difference in the production of infectious herpes simplex virus. Taken together, these findings suggest that when commercially-useful cells such as normal fibroblasts and epithelial cells are grown in large quantities on microcarriers, the nature of the substrate may have a profound effect on the growth and physiology of the cells. They also suggest that when microcarriers are used, unexpected results based on preliminary work in conventional monolayer culture may be obtained.
Varani
J
J
Dame
M
M
Rediske
J
J
Beals
T F
TF
Hillegas
W
W
D000818:Animals; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003584:Cytological Techniques / Q000295:instrumentation; D003637:DEAE-Dextran; D004847:Epithelial Cells; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism / Q000382:microbiology; D006801:Humans; D008855:Microscopy, Electron, Scanning; D010447:Peptide Hydrolases / Q000378:metabolism; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D011453:Prostaglandins / Q000096:biosynthesis; D018139:Simplexvirus / Q000254:growth & development; D013379:Substrate Specificity; D013997:Time Factors; D014357:Trypsin / Q000494:pharmacology
D011453:Prostaglandins; D003637:DEAE-Dextran; D011451:Prostaglandin-Endoperoxide Synthases; D010447:Peptide Hydrolases; D014357:Trypsin