A系小鼠胚腭突细胞培养及生物学特性的研究

A Study of the Combined Culture of A/J Mouse Embryonic Palatal Mesenchymal and Epithelial Cells andTheir Biological Behavior in Vitro

目的:研究A系小鼠胚腭突上皮与问充质细胞在DMEM/F-12培养基中进行原代和传代联合培养及其生物学特性。方法:采用机械分离及胰蛋白酶消化法,简便、准确、快速地获得高成活率的A系小鼠胚腭突细胞,并通过流式细胞仪、相差显微镜、光镜等观察其DNA及蛋白质合成、细胞骨架及其结构。结果:联合培养的A系小鼠胚腭突细胞中,腭突上皮及问充质细胞均能较好地生长,光镜下观察见上皮型细胞呈多角型,单层铺路石状排列,细胞骨架为栅栏型或巢穴型;间充质细胞呈梭型,单层漩涡状排列,细胞骨架呈梭型、多平行排列的纤维。流式细胞仪检测见细胞DNA及蛋白质含量较丰富,细胞增殖活跃。结论:A系小鼠胚腭突上皮与间充质细胞联合培养模型能较好地保持体内胚腭突的上皮细胞及问充质细胞细胞成分的基本特征。

Objective: The aim of this study was to establish a combined culture model of A/J mouse embryonic palatal shelves mesen-chymal and epithelial cells (palatal cells) , and study their biological behavior in vitro.Methods: The A/J mouse embryonic palatal cells were harvested from 13-day-old A/J mouse fetus by dissection and sequential digestion with 0.25% trypsin. The isolated cells were cultured in DMEM/F-12 medium and observed by phase-contrast microscope, flow cytometer, cytoskeleton staining and PGEj-RIA. Results: The embryonic palatal mesenchymal and epithelial cells grew well and, their morphology and cytoskeleton did not change in vitro. The cells were full of abundant protein and DNA, and had strong ability to metabolize and proliferate. Conclusion: The mesenchymal and epithelial cells of A/J mouse embryonic palatal shelves in this combined culture system can keep their basic biological behavior.