大鼠羊膜上皮细胞的生物学特性  

The biological characteristics of amniotic epithelial cells derived from rat amnion

作　　者：段现花[1] 王德广[1] 欧阳长杰[1] 曲德伟[1] 

机构地区：[1]徐州医学院解剖学教研室,江苏徐州221002

出　　处：《中国老年学杂志》2013年第9期2068-2070,共3页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(No.31271358);徐州市科技发展基金(No.XF10C079);徐州医学院院长专项人才基金(No.2011KJZ25)

摘　　要：目的探讨建立大鼠羊膜上皮细胞分离、培养及鉴定方法及观察其生物学特性。方法从妊娠14～16 d的SD大鼠胎盘剥离羊膜,用胰蛋白酶消化法分离大鼠羊膜上皮细胞,采用免疫荧光细胞化学和流式细胞仪分析细胞表型特征;使用含有10%胎牛血清(FBS)和100 U/ml青霉素-链霉素(PS)溶液的DMEM/F-12的培养基进行细胞培养。结果每次采用胰蛋白酶消化法从1只孕鼠分离的上皮细胞数为(1～6)×105/ml,足够达到贴壁培养浓度。所分离的羊膜上皮细胞表达神经前体细胞特异性蛋白nestin和骨髓间充质干细胞表面标记蛋白CD29和CD44,但是未成熟神经元标记蛋白β-Ⅲ-tublin、星形胶质细胞标记物胶质纤维酸性蛋白(GFAP)以及造血干细胞表面标记蛋白CD34表达阴性。在完全培养基培养条件下,羊膜上皮细胞生长增殖较快,原代培养1 w后即可传代。结论建立了大鼠羊膜上皮细胞分离、培养及鉴定的方法,大鼠羊膜上皮细胞具有神经前体细胞的特异标志和间充质干细胞的某些表型特征。Objective To investigate the isolatioH aud culture condition of rat anmiotic epithelial cells （rAECs） and its phenotypic identification, and observe their biologitcal characteristics. Methods rAECs were isolated from pregnancy 14 - 16 d SD rat anmion digested with 0. 25% trypisin. The phenotype of rAECs were identified by immunocytochemistl7 and Flow Cytometry. These cells were cultured in DMED/F12 medium contained 10% FBS and 100 U/ml PS. Results The amnion originated from a rat was obtained （1 - 6 ） × 10^5/ml rAECs by trypisin digestion. Nestin, as a neural progenitor cell umrker, was expressed in the rAECs derived from rat amnion, and mesenchymal stem cells surface marker CI）29 and CIM4 extendly different exteudly were expressed, while β-III-tublin protein and GFAP, respectively as marker for immature neurons and astrocyle, was negalive, and hemalopoielitic stem cells marker CD34 were not detected. The total cell number rAECs, cultured fur 7 d after primary, were expanded hy subcultured. Conclusions A method for isolation, cultured and identific, ation of rAECs derived from rat amnion are successfully established. The rAECs have specific marker of neural system cells and some phenotypic properties of mesenchymal stem cells.

关 键 词：大鼠羊膜上皮细胞 分离 培养 鉴定 

分 类 号：R74[医药卫生—神经病学与精神病学]