大鼠腹膜间皮细胞的分离培养方法  

Establishment of method for primary culture and passage of rat peritoneal mesothelial cells

作　　者：马姝琛[1] 严海东[1] 庄守纲[1] 兰洋[1] 张瑞青[1] 王奕[1] 

机构地区：[1]同济大学附属东方医院肾内科,上海200120

出　　处：《同济大学学报（医学版）》2012年第6期19-21,共3页Journal of Tongji University(Medical Science)

基　　金：国家自然科学基金(81170638)

摘　　要：目的建立简便有效的大鼠腹膜间皮细胞(passage of rat peritoneal mesothelial cells,RPMC)的体外培养方法。方法采用0.25%胰蛋白酶-0.02%EDTA消化大鼠大网膜及脾胃韧带,细胞悬液进行离心培养,沉淀重悬后进行培养。通过形态学、免疫荧光、免疫组化法鉴定细胞。结果分离培养的细胞在光镜下呈铺路鹅卵石状,免疫荧光鉴定显示细胞角蛋白阳性,免疫组化鉴定显示波形蛋白抗原阳性,第Ⅷ因子、白细胞CD45抗原阴性,证实培养的细胞为RPMC。结论胰蛋白酶消化法是一种简便而实用的分离RPMC的方法。Objective To establish a convenient and effective method for primary culture and passage of rat peritoneal mesothelial cells（RPMCs）.Methods Omental specimens were obtained from SD rats and digested with 0.25% trysin-0.02%EDTA for 10 min.Cell deposit was suspended and was cultured in DMEM/F12 containing 15% fatal calf serum.Cells were verified by morphology,cytoimmunohistochemistry and cytoimmunofluorescence analysis.Results The confluent cells showed cobblestone-like appearance under microscope.Cytoimmunofluorescence analysis showed a positive staining of cytokeratin.Cytoimmunohistochemistry analysis showed a positive staining of vimentin,a negative staining of CD45 and factor Ⅷ.Conclusion The method established in the study is an easy and practical one to obtain RPMCs.

关 键 词：腹膜间皮细胞 原代培养 大网膜 大鼠 

分 类 号：R331[医药卫生—人体生理学]