大鼠口腔黏膜固有层前体细胞与外胚间充质细胞生长特性的研究  

Separation and growth proliferation characteristics of oral mucosal lamina propria progenitor cell and ecto-mesenchymal stem cells

作　　者：董蕊[1] 刘晓亮 葛丽华[1] 樊明文 

机构地区：[1]首都医科大学口腔医学院口腔医学研究所,北京100050 [2]武汉大学口腔医学院口腔生物医学工程教育部重点实验室

出　　处：《北京口腔医学》2012年第6期305-309,共5页Beijing Journal of Stomatology

基　　金：国家自然科学基金(30700946);北京市自然科学基金(7112057);北京市科技新星计划(2011083)

摘　　要：目的比较大鼠口腔黏膜固有层前体细胞(oral mucosal lamina propria progenitor cell,OMLPPC)与面突外胚间充质细胞(ecto-mesenchymal stem cells,EMSC)体外生长特性。方法免疫磁珠筛选OMLPPC,差速贴壁法分离EMSC。利用免疫组织化学方法对分离细胞进行鉴定。分别利用CCK-8、流式细胞等方法检测其生长增殖能力、细胞周期,并检测细胞克隆形成能力,观察并比较两种细胞的体外增殖特性。结果体外培养过程中,两者都是梭形类成纤维样细胞,OMLPPCs贴壁、克隆样生长,细胞汇合后,细胞呈短梭形。生长曲线结果显示,OMLPPCs无明显的生长滞缓期,8天后仍保持强的增殖活力。EMSCs具有明显的缓慢生长期、对数生长期及平台期。细胞克隆形成能力比较,EMSCs的克隆形成能力比OMLPPCs强。流式细胞检测结果显示,EMSCs第1代增殖指数为9.78,OMLPPCs为14.97;第2代时EMSCs为40.04,OMLPPCs为30.38。结论 OMLPPC具有与EMSC相似的间充质干细胞形态,EMSCs为更早胚胎组织来源的干细胞,而OMLPPCs细胞组成相对单一。OMLPPCs在体外生长过程中具有较强的增殖适应性,可以作为间充质前体细胞的来源,为相关口腔组织的组织工程构建提供候选种子细胞。Objective To compare the proliferation characteristics of rat oral mucosal lamina propria progenitor cell （OMLPPC） with ecto-mesenchymal stem cells （EMSC） in vitro. Methods Rat OMLPPCs were isolated by an immunomagnetic sorting technique, and EMSCs by enzyme digestion, which were identified by immunocytochemistry respectively. The growth proliferation was observed and analyzed by CCK-8, flow cytometer （FCM） and colony-forming unit assay. Results The morphological characteristics of OMLPPCs were similar to those of EMSCs. Both looked like the mesenchymal cells. But the shape of OMLPPCs was uniform and showed short fusiform. EMSCs had standardized cell growth curve, but the OMLPPCs had high viability in the 8th day. The cloning efficiency of EMSCs was higher than that of OMLPPCs. From the first passage to the second , the proliferation index of OMLPPCs and EMSCs increased, which was from 9.78 to 40. 04 in EMSCs, and from 14. 97 to 30. 38 in OMLPPCs. Conclusion Both EMSCs and OMLPPCs looked like the mesenchymal stem cells. The proliferation ability of OMLPPCs was similar to that of EMSCs. OMLPPCs may serve as a suitable cell source for future tooth tissue engineering strategies.

关 键 词：口腔黏膜固有层 前体细胞 外胚间充质细胞 增殖 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]