人胎盘滋养层细胞的分离培养及IgG FcγRⅢ在滋养层细胞的表达  

Isolation of villous cytotrophoblast from term human placenta and expression and location of IgG FcγRⅢ in human term placenta

作　　者：程勇前[1] 聂青和[1] 周永兴[1] 杜德伟[1] 杨华光[2] 

机构地区：[1]第四军医大学唐都医院解放军感染病诊疗中心,陕西西安710038 [2]第四军医大学唐都医院妇产科,陕西西安710038

出　　处：《医学研究生学报》2002年第2期105-108,111,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金资助项目 (编号 39970 767)

摘　　要：目的 :体外分离培养人胎盘滋养层细胞 ,观察其生物学特性 ,研究IgGFcγRⅢ (CD16)在人胎盘组织中的定位及体外培养的滋养层细胞是否存在CD16,从而为HCV母婴传播机制的研究提供细胞学实验基础。　方法 :采用胰蛋白酶消化法消化人足月胎盘组织 ,以 3 5 %、4 5 %两个Percoll密度梯度进行分离纯化。用ABC免疫组化染色法对足月分娩后的胎盘组织石蜡包埋切片及体外分离培养的胎盘滋养层细胞进行染色。　结果 :胎盘组织中滋养层细胞角蛋白染色阳性 ,血管内皮细胞及基质成分波形蛋白染色阳性 ,经该法分离纯化的细胞角蛋白染色阳性者(滋养层细胞 )占 90 %以上 ,胎盘组织切片的滋养层细胞及体外分离培养的滋养层细胞抗CD16染色阳性 ,阳性信号定位于胞质 ,未见胞核着色。　结论 :改进后的分离方法可以得到较高纯度的滋养层细胞 ,胎盘组织的滋养层细胞和体外分离培养的滋养层细胞均有CD16表达。Objectives: To isolate cytotrophoblasts from term human placentae, observe their biological characteristics and study the location of CD16 in human placenta to see whether there is CD16 in the cytotrophoblasts cultured in vitro , and provide a basis for cellular experiment of mother to infant transmission of HCV. Methods:Cytotrophoblasts were prepared from term human placentae digested with trypsin, and centrifuged on two layers(35% and 45%) of Percoll gradient. Paraffin sections of placenta and trophoblasts cultured in vitro were stained with ABC immunohistochemistry staining. Results: In human placentae, the trophoblast layer was positive for cytokeratin and negative for vimentin, while endothelial cells of blood vessel and hypothalamus were negative for cytokeratin and positive for vimentin. More than 90% cytokeratin positive cells (trophoblast) were obtained from our improved method. Positive stain was observed on trophoblasts in the placenta sections and those cultured in vitro , and the positive signal was located in the cytoplasm, not in the nucleus. Conclusions: High purity trophoblasts could be obtained by this improved method. CD16 existed in the trophoblasts in the placenta and trophoblasts cultured in vitro .

关 键 词：滋养层细胞 分离纯化 细胞培养 FC受体 体外实验 表达 

分 类 号：R329-33[医药卫生—人体解剖和组织胚胎学]