人胃黏膜成纤维细胞原代培养条件的优化  

Optimal primary culture conditions of human gastric mucosal fibroblasts

作　　者：张浩[1] 申晓军[1] 唐古生[2] 毕建威[1] 

机构地区：[1]第二军医大学附属长海医院普外科,上海200433 [2]第二军医大学附属长海医院检验科,上海200433

出　　处：《中华实验外科杂志》2009年第5期610-611,共2页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基会资助项目（30672046）

摘　　要：目的通过优化人胃黏膜成纤维细胞的原代培养条件，提高培养成功率，为进一步研究胃癌相关成纤维细胞奠定基础。方法原代培养采用植块法，利用免疫组织化学、细胞形态学和电镜等方法进行细胞鉴定；分析不同培养表面、培养基和培养液pH值对原代细胞培养游出情况的影响，筛选适合的培养条件。结果Logistic逐步回归分析提示RPMI1640培养基的WaldΧ^2值=32．4533，P〈0．01，标准化回归系数=0．7852，说明使用RPMI1640培养基更利于原代成纤维细胞游出；pH=7．4的培养液的WaldΧ^2值：49．4756，P〈0．01，标准化回归系数=0．4867，说明培养液pH值为7．4更利于原代成纤维细胞游出；不同水平的培养表面条件对成纤维细胞游出率的影响差异无统计学意义（P〉0．05），而且各培养条件之间无交互作用。结论通过优化培养表面、培养基种类和pH值等培养条件，可以提高人胃黏膜成纤维细胞原代培养的成功率。Objective To optimize primary culture conditions of human gastric mucosal fibroblasts for further investigations of gastric cancer associated fibroblasts. Methods The fibroblasts were cultured primarily by explant method, and identified by immunohistochemistry, morphology and electron microscopy. The effects of different culture conditions on emigration ratios of cells were investigated to optimize the culture surface, medium and pH in primary culture of gastric mueosal fibroblasts. Results When the other conditions were the same, no significantly different emigration rate was found on the mouse tailcoated surface, fetal bovine serum-coated surface, and nothing coated surface （ P 〉 0.05 ）. As to the role of different culture media including RPMI 1640 medium, DMEM and high-gluco DMEM, there was significantly higher emigration rate in the RPMI 1640 medium compared with other two media （P 〈 0.01, standardized estimate = 0. 4688 ）. Different pH values （ pH = 7.0,7.4 and 7.8 ） of culture solution led to different emigration rate, and the best emigration rate was found in pH = 7.4 solution （P 〈 0. 01, standardized estimate = 0.5857 ）. Furthermore, there was no interactional effect on the cell emigration rate among culture conditions. Conclusion The primary culture of human gastric mueosal fibroblasts can be greatly improved by optimizing culture conditions such as kinds of medium and pH values of culture solution.

关 键 词：胃黏膜 成纤维细胞 原代培养 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]