[Vital long-term preservation of human gingiva in perfusion culture]

Perfusion culture offers the advantage of keeping gingiva alive for a long time as an stable explant according to cell biological parameters. To investigate the suitability of cultured human gingival explants for transplantations the biopsies were put into a newly developed perfusion chamber and cultured for at least 21 days. Gingiva explants were derived from healthy donors undergoing surgical removal of molar teeth. The tissue pieces were cultured without prior proteolytic desintegration or subculture. Immediately after excision a morphological and immunohistochemical analysis of the tissue was carried out and the distribution pattern of cytokeratin and vimentin was examined. Gingival explants cultured for 7, 14 and 21 days in serum-free keratinocyte growth medium in perfusion culture were analyzed in the same way. The morphology of the cultured explant (21 days) was well preserved from stratum basale up to stratum corneum. As proved by immunohistochemical incubation with antibodies to CK 5/6, CK 14 and CK 19, a tissue-specific cytokeratin (CK) expression pattern was maintained during the whole perfusion period. After 7 days of culture vimentin was synthesized in the fibroblast layer and was found in small quantities in each layer of the epithelium. In contrast to conventional cultures, where dissociation of the tissue and a subculture interruption is usually needed for long-term culture, this is not necessary for perfusion cultured tissue. The use of perfusion-cultured gingival explants as autogenous transplants is investigated herein.