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Establishment of an osseous cell line from fetal rat calvaria using an immunocytolytic method of cell selection: Characterization of the cell line and of derived clones
Author(s) -
Bernier Suzanne M.,
Desjardins Johanne,
Sullivan Arthur K.,
Goltzman David
Publication year - 1990
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041450212
Subject(s) - alkaline phosphatase , cell culture , endocrinology , medicine , retinoic acid , population , parathyroid hormone , biology , osteocalcin , osteoblast , cellular differentiation , chemistry , microbiology and biotechnology , biochemistry , in vitro , calcium , enzyme , genetics , environmental health , gene
Abstract Using selective media and complement‐mediated lysis of primary cultures of a fetal rat calvarial cell population, we have developed a cell line (OBCK6) that exhibits osteoblastic characteristics. OBCK6 cells demonstrated enhanced para‐thyroid hormone (PTH)‐stimulated adenylate cyclase activity relative to the primary, calvarial population, production of alkaline phosphatase activity and type I collagen, and the capacity to form mineralized nodules in unsupplemented medium after prolonged (22–26 day) culture. Two sublines, CFK1 and CFK2, which were isolated by dilution cloning, differed morphologically and with respect to growth rate. CFK1 cells demonstrated high PTH and prostaglandin E 2 ‐stimulated adenylate cyclase activity, whereas only low PTH‐stimulated activity was observed in CFK2 cells. Retinoic acid and 1,25‐dihydroxyvitamin D 3 [1,25(OH) 2 D 3 ] each reduced PTH‐stimulated adenylate cyclase activity in both the cell types. Retinoic acid and dexamethasone reduced and 1,25(OH) 2 D 3 enhanced alkaline phosphatase activity in these cells. PTH significantly augmented alkaline phosphatase activity to a much greater extent in CFK1 than in CFK2 cells. Both CFK1 and CFK2 cells expressed type I but not type III collagen, and neither expressed osteocalcin. Strong Alcian blue staining of CFK2 cells was suggestive of a cartilaginous phenotype. These three cell lines, therefore, demonstrated discrete characteristics of skeletal cell function and should provide important models for evaluation of mechanisms of mineralization and for control of skeletal cell growth and mesenchymal differentiation in vitro.