第一鳃弓外胚间充质细胞体外培养模型的构建☆

背景：在牙颌面的发育形成过程中，第一鳃弓作为过渡结构，发挥着不可替代的重要作用。目前，国内外对于第一鳃弓外胚间充质细胞的研究资料相对较少。 目的：分离培养小鼠第一鳃弓外胚间充质细胞。 方法：显微解剖E9.5胎鼠第一鳃弓，采用组织块法和酶消化法对其进行原代培养，通过细胞形态观察、细胞生长曲线描记、计算细胞倍增时间和特异性标记物检测对其细胞生物学特性进行研究。 结果与结论：组织块法原代培养细胞的时间7~10 d，上皮细胞成分较多，酶消化法原代培养细胞时间两三天，上皮细胞成分较少，两组细胞传代后细胞形态、细胞生长曲线、群体倍增时间无明显差异。特异性标记物CD57/HNK1、波形丝蛋白免疫细胞化学染色呈均一表达。提示采用酶消化法可以在短时间内获得数量充足、纯度较高、满足实验要求的细胞。

Establishment of an in vitro culture system of ectomesenchymal cells isolated from the first branchial arch

BACKGROUND: During the dentofacial development, the first branchial arch as a transitional structure plays an important role. At present, domestic and international research data for the first branchial arch mesenchymal cells were relatively few. OBJECTIVE: To isolate and culture the mesenchyme cells from the first branchial arch of the mice embryos. METHODS: The first branchial arch primordia from E9.5 embryos were dissected under a microscope. Explanted culture and improved enzyme digestion were used to primarily culture. The growth curve, population doubling time, cells proliferation and morphology were studied and specific markers were detected to study biological characteristics of cells. RESULTS AND CONCLUSION: The cells were grown to confluency for 7-10 days, with abundant epithelial cells. Improved enzyme digestion of primary culture had less epithelial-like cells, the cells were grown to confluency for 2-3 days. The growth curve, population doubling time and morphology were similar in both groups after passage. Anti-CD57/HNK-1 and anti-vimentin were all positive. Results have indicated that sufficient and high purity ectomesenchymal cells could be harvested in short time by improved enzyme digestion.