Characterization of Cultured Rat Embryonic Palatal Mesenchymal Cells

Abstract

To investigate developmental palatogenesis, the establishment of palatal cell culture in vitro is preferable to eliminate several complicated biases present in the in vivo environment. We established a primary culture of rat embryonic palatal mesenchymal cells using a special technique to dissect embryonic palatal shelves, and characterized these embryonic cells by immunohistochemical analysis against histiocytic markers. Following preparation of the maxilla of 15.5-day-old rat fetuses, a midline incision of the maxilla was established while the occiput was fixed with microforceps. This procedure allowed eversion of the maxillary process and easy dissection of the palatal shelf. The technique allowed preparation of a large number of palatal shelves with no appendages using a small number of fetuses. Cells cultured with DMEM/F-12 and 10% FBS showed multi-potential nature (i.e., not only mere mesenchymal character but also neural, endothelioid, and/or myoblastoid origin were identified by immunostaining with anti-epithelium membrane antigen, keratin, vimentin, S-100 protein, factor VIII, desmin, and lysozyme antibodies, respectively). Our results demonstrated that, during several cell passages, the cultured cell gained myoblastoid characteristics in addition to a neural nature. Further in vitro studies using cultured embryonic palatal mesenchymal cells will assist in characterization of proliferation and differentiation of cells forming the palate.