Tendons from 14-17-day-old chick embryos contain predominantly type I collagen and about S o / , AB2 collagen; type 111 collagen is not detectable by biochemical methods, such as sodium dodecyl sulfate/polyacrylamide gel electrophoresis or cyanogen bromide pattern, but can be visualized by immunofluorescence staining with collagen-type-specific antibodies.Similarly, freshly dissociated tendon cells secrete only type I collagen into the culture medium but no significant amounts of type I11 collagen [Uitto, J., Lichtenstein, J. R., and Bauer, E. A. (1976) Biochemistry, 15, 4935-49421. Transfer of tendon cells from chick embryos to monolayer conditions, however, initiated synthesis of type I11 collagen in about 10% of the cells within three days, as visualized by immunofluorescence staining. Secretion of type I11 collagen into the culture medium can also be detected by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. With increasing number of passages the number of cells producing type 111 collagen reached levels of about 80 % after the third passage, while 90% of all cells stained positively for type I collagen. This is reflected by an increase of production of type 111 collagen as determined by CM-cellulose chromatography. Using velocity sedimentation, the secretion of type 111 procollagen and of pN-collagen (carrying the aminoterminal extension only), into the culture medium of a second-passage tendon cell culture was detected.This study provides new evidence that the phenotype of cells may alter during transfer from the environment in vivo to conditions in vitro and that additional changes may occur with time in culture.Fibroblasts from various sources show different, tissue-specific features such as size, cell shape [l] and cell surface antigens [2]. They also differ in the relative proportions of proteoglycans and collagen types synthesized. Most fibroblasts, such as skin fibroblasts [3,4], rabbit lung fibroblasts NB6 [5] and several other established cell lines [6], produce both types I and I11 collagen in different ratios, with a preponderance of type I collagen. However, there are also fibroblasts which produce only type I collagen, e.g. chick cornea fibroblasts [7].It is generally believed that the pattern and relative proportions of collagen types synthesized by cells in vitro reflect the collagen composition of the tissue from which the cells originated. For example, the Other cells, however, may change the ratios of collagen type synthesis when released from tissues and transferred to systems in vitro ; also the qualitative pattern of collagen type synthesis may be altered. For example, chondrocytes change from synthesis of type TI collagen in vivo to synthesis of types I and I11 collagen under certain conditions in vitro [ l l -131. Embryonic chick cornea fibroblasts, which produce only type I collagen in vivo [7], immediately begin to synthesize type 111 collagen in addition to type I collagen after dissociation of the corneal matrix [14].On the other hand, cultures of established cell l...