Development of a new serum-free medium,USC-HC1, for growth and normal phenotype in postembryonic chicken growth plate chondrocytes |
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Authors: | Laura V Hale John E Hale Mary Lynn S Kemick Yoshinori Ishikawa Roy E Wuthier |
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Institution: | (1) Department of Chemistry, University of South Carolina, 29208 Columbia, South Carolina |
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Abstract: | Summary A serum-free medium for postembryonic chicken epiphyseal growth plate chondrocytes has been developed from 104 MCDB medium.
To enable these fastidious cells to survive, grow, and express normal phenotype, a substantial increase over MCDB 104 in the
level of many of the amino acids was required, as well as a change in the buffer system and the addition of SerXtend, a defined,
serum-free product containing various growth factors, including fibroblast growth factor. Also required was the provision
of cell attachment factors, either by coating culture surfaces with type II collagen, or better, by allowing the freshly released
cells to recover for several hours in a medium supplemented with 10% fetal bovine serum before plating. Ths new serum-free
medium, which we call USC-HC1, supports growth and replication, the retention of normal polygonal morphology, the expression
of significant levels of cellular alkaline phosphatase activity, the production of sulfated proteoglycans, type II collagen,
and the formation of alkaline phosphatase-rich matrix vesicles by the chondrocytes. The major advantage of USC-HC1, however,
is that it will provide for the first time an opportunity to examine the effects of various defined growth and hormonal factors
on the phenotypic expression and differentiation of growth plate chondrocytes, in the absence of the variable (stimulatory
and inhibitory) factors present in fetal bovine serum.
This work was supported by grant AM18983 from the National Institute of Arthritis, Diabetes, Digestive and Kidney Diseases,
Bethesda, MD. |
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Keywords: | growth plate chondrocytes serum-free medium primary cell culture matrix vesicles alkaline phosphates |
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