A quantitative assay for collagen synthesis in microwell fibroblast cultures.

A new method for estimating collagen synthesis in microwell cultures of fibroblasts is presented, ³H-Labeled-proline-collagen was purified by successive salt precipitations at acid and neutral pH in the presence of carrier collagen. Variability between replicates was less than 10% (standard deviation) and recovery of labeled collage internal standards was greater than 90%. More than 90% of recoverable radioactivity was in collagen as demonstrated by polyacrylamide gel electrophoresis and carboxymethyl cellulose chromatography. The culture system is highly reproducible and allows use of a large number of separate cultures with uniformity of culture conditions and economy of reagents.