大鼠小肠上皮细胞的体外原代培养  

In vitro primary culture of rat intestinal epithelial cells

作　　者：王莉[1] 段相林[1] 

机构地区：[1]河北师范大学生命科学学院,河北石家庄050016

出　　处：《军事医学科学院院刊》2004年第1期61-63,共3页Bulletin of the Academy of Military Medical Sciences

基　　金：河北省自然科学基金 ( 3 0 3 15 8) ;河北省教育厅自然基金项目 ( 2 0 0 2 13 6)

摘　　要：目的 :探讨大鼠小肠上皮细胞原代培养的原理与方法以及培养过程中常见的问题和克服办法。方法 :取生后 1周SD大鼠的小肠上皮细胞 ,经分离纯化后进行原代培养。结果与结论 :胶原酶Ⅺ型和中性蛋白酶Ⅰ型配合使用 ,可以分离到形态完整、容易贴壁生长的小肠绒毛细胞团。进一步利用离心技术纯化得到 95 %以上的小肠上皮细胞。结果显示 ,常规体外培养的肠上皮细胞是否增殖分化最终决定于用酶消化后的结构完整性 ,如果用0 .2 5 %胰酶消化 ,则细胞受损程度较大 ,不易贴壁生长。Objective:To explore the principle and method for growing small intestinal epithelial cells in short-term primary cultures and some commonly encountered problems during the culture process in order to resolve them. Methods:The intestinal epithelial cells were taken from postnatal 1 st week Sprague-Dawley(SD) rat for primary culture after being separated and purified. Results and Conclusions:Isolation of the epithelial cells and preservation of their three-dimensional integrity were achieved using the digestion technique with a mixture of collagenase Ⅺ and dispaseⅠ. Purification of the epithelial cells was facilitated by the use of a simple differential sedimentation method. The results show that proliferation of normal gut epithelium in vitro is initially dependent upon the maintenance of the structural integrity of the tissue. If the 0.25% trypsin is used for digestion, the cells would be severely harmed and are very difficult to stick to the Petri dish for growing.

关 键 词：小肠 上皮细胞 体外培养 大鼠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]