Characterizing Pluripotency of Primary Cells Derived from Elasmoid Scales of Zebrafish (Danio rerio)

Recently, an in vitro tooth organ culture system was developed using pluripotent stem cells from rat incisors. This model system recapitulates the reciprocal epithelial-mesenchymal interactions exhibited by many organs during embryogenesis. Remarkably, the tooth organ culture system gave rise to a tooth, that when transplanted, regained most of the necessary attributes of ontogenic rat incisors. However, noticeable decreases in dental type protein expression levels were identified in the in vitro tooth organ. The issue with the protein levels will need to be resolved in order to develop an accurate tooth replacement organ. It has been suggested that elasmoid scales are derived from odontodal tissues covering ancestral osteichthyan rhombic scales. Elasmoid scales of zebrafish express nearly all the same genes known for contributing to tooth patterning in mammals. These scales also exhibit similar developmental phases including induction through reciprocal epithelial-mesenchymal interactions. Developing an in vitro elasmoid scale organ culture system would be beneficial for studying processes to up-regulate protein expression during the induction of reciprocal epithelial-mesenchymal interactions. Therefore, the aim of this research is to isolate mesenchymal and ectodermal cells from zebrafish scales and characterize their pluripotency as potential candidates for use in a scale organ culture system. Thus, isolated cells were exposed to a variety of growth media to influence differentiation. These cell lines were then analyzed by RT-PCR for osteoblast, epithelial, and nervous tissue cellular markers to measure the differentiation potency