[Ultrastructural and cytochemical studies of clonal cell lines in fibroblasts derived from human periodontal ligaments--establishment of clonal cell lines in fibroblasts from periodontal ligaments].

Abstract

It has recently been suggested that fibroblasts in periodontal ligaments have cytochemical characteristics similar to those of osteoblasts and different from those of other connective tissues. The authors isolated clonal cell lines of fibroblast-like cells from human periodontal ligaments in order to clarify their nature. Digestion with collagenase and hyaluronidase was used to isolate the cells from a human periodontal ligament. The cells were then plated in a 96-hole microplate. A single cell in a conditioned medium containing 20%FBS was placed in each hole. From these single cells large colonies ware subcultured. Subculturing was done every 8 days until more than 20 successive generations had been produced. The method developed by Lowry et al. was used to determine the ALPase activity of the cultured cells. From the 768 cells cultured from human periodontal ligament, 7 clonal cell lines were isolated in vitro. Cultures of these clonal cell lines resulted in typical, spindle-shaped, fibroblast-like cells, all of which were homogeneous. Very high ALPase activity was observed in 4 of the 7 cell lines. Enzyme reaction products occurred mainly along cell membranes. These stable clonal cell lines provide suitable systems for in vitro studies related to morphological and functional analysis of fibroblasts in the periodontal ligament.