Anin vitro, serum-free organ culture technique for the study of development and growth of the dermal skeleton in fish

Abstract

To develop a serum-free, chemically definedin vitro organ culture system enabling the study of epithelial-mesenchymal interactions in development and growth of fish dermal skeleton, we investigatedin vitro continuation of scale regeneration in the cichlid fishHemichromis bimaculatus. The culture medium in our system is based on Leibovitz medium (L-15) supplemented with vitamin C, additional amino acids and HEPES. With this basis medium, we examined the effects of all trans-retinoic acid, dexamethasone, and prostaglandin-E2 (PG-E2), factors known to exert an effect on development and growth of teeth and bone in mammalian culture systems, on thein vitro regeneration of scales. These effects were compared with those obtained by supplementation of the basis medium with newborn and fetal calf serum. To evaluate our culture system, the medium that allowed to mimick in the best possible way thein vivo regeneration of scales (i.e., the basis medium plus dexamethasone and PG-E2) was also tested on thein vitro development of teeth in the same fish species.