Serum-free culture of rabbit meniscal fibrochondrocytes: proliferative response.
We have formulated a serum-free medium capable of supporting DNA synthesis in rabbit meniscal fibrochondrocytes at a level equivalent to 10% fetal bovine serum (FBS). The medium consists of a 1:1 mixture of Dulbecco's Modified Eagle's Medium and Ham's F-12 medium supplemented with transferrin (1 microgram/ml), selenium (1 pg/ml), trace metal mix (1:100), dexamethasone (100 ng/ml), insulin-like growth factors I and II (50 ng/ml each), pituitary fibroblast growth factor (100 ng/ml), and lactalbumin hydrolysate (2 micrograms/ml). Endothelial cell growth supplement could be substituted for lactalbumin hydrolysate to obtain similar results. Ventrex PC-1, a commercially available, low-protein, serum-free medium, was found to support proliferation of fibrochondrocytes but not as well as 10% FBS or our medium formulation. Lipid supplements, which are known to support the serum-free growth of hyaline chondrocytes, were found to be either of no value or antagonistic for the culture of fibrochondrocytes. Likewise, vitamin E alone, progesterone, putrescine, and hydrocortisone were also without benefit in our culture system. The cells had a more chondrocytic morphology when grown in defined medium as opposed to 10% FBS. The results of this study should now make it possible to identify and quantitate those factors necessary to affect meniscal repair by utilizing further techniques in vitro.
Webber
R J
RJ
Department of Pathology, University of Arkansas for Medical Sciences, Little Rock 72205.
Zitaglio
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Hough
A J
AJ
D000818:Animals; D002417:Cattle; D002455:Cell Division; D002478:Cells, Cultured; D003470:Culture Media; D004261:DNA Replication; D005312:Fetal Blood; D006133:Growth Substances / Q000494:pharmacology; D008592:Menisci, Tibial / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D011817:Rabbits; D013936:Thymidine / Q000378:metabolism; D014316:Tritium
D003470:Culture Media; D006133:Growth Substances; D014316:Tritium; D013936:Thymidine