Yano H, Yoshimoto H, Ohtsuru A, Ito M, Yamashita S, Fujii T
Department of Plastic and Reconstructive Surgery, Nagasaki University School of Medicine, Japan.
Cleft Palate Craniofac J. 1996 Sep;33(5):379-84. doi: 10.1597/1545-1569_1996_033_0379_cocrep_2.3.co_2.
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 multipotential 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.
为了研究发育性腭形成，建立腭细胞体外培养体系以消除体内环境中存在的多种复杂偏差是较为可取的。我们采用一种特殊技术解剖胚胎腭突，建立了大鼠胚胎腭间充质细胞的原代培养体系，并通过针对组织细胞标志物的免疫组织化学分析对这些胚胎细胞进行了表征。在制备15.5日龄大鼠胎儿的上颌骨后，在固定枕骨的同时对上颌骨进行中线切口。该操作使上颌突外翻，便于腭突的解剖。该技术能够使用少量胎儿制备大量无附属物的腭突。用DMEM/F-12和10%胎牛血清培养的细胞表现出多能性（即，通过分别用抗上皮膜抗原、角蛋白、波形蛋白、S-100蛋白、因子VIII、结蛋白和溶菌酶抗体进行免疫染色，不仅鉴定出单纯的间充质特征，还鉴定出神经、内皮样和/或成肌样起源）。我们的结果表明，在几次传代过程中，培养的细胞除了具有神经特性外，还获得了成肌样特征。使用培养的胚胎腭间充质细胞进行进一步的体外研究将有助于表征形成腭部的细胞的增殖和分化。
