人胎盘滋养层细胞的分离和鉴定

Isolation and identification of human placenta cytotrophoblast

目的:体外分离足月妊娠胎盘滋养层细胞,观察其形态学特点,检测人类白细胞抗原-E(HLA-E)在滋养层细胞的表达情况。方法:采用0.25%胰酶和胶原酶I联合消化法及Percoll密度梯度分离法分离人足月妊娠胎盘组织中滋养层细胞,光学显微镜观察分离出的细胞形态,透射电镜观察分离出的细胞超微结构;RT-PCR技术检测分离出的滋养层细胞HLA-E的表达。结果:光学显微镜下初分离的细胞呈大圆形,以单个核为主;透射电镜下可见细胞表面的微绒毛丰富,胞质丰富,内质网扩张明显,有丰富的糖原颗粒、髓样小体和明显的脂滴。在分离的细胞上检测到HLA-E的表达。结论:采用胰酶和胶原酶I联合消化及Percoll密度梯度分离法可获得高纯度的滋养细胞,在透射电镜下可见到超微结构,同时检测到滋养层细胞上表达HLA-E。

Objective： To isolate cytotrophoblasts from term human placenta, observe their morphology characteristic. To detect the expression pattern of human leukocyte antigen HLA - E mRNA in normal human third trimester placenta. Methods： Cytotrophoblasts were prepared from term human placenta digested with trypsin, collagenase 1 and density gradient separation, to evaluate amorphous and formation of cytotrophoblast by light microscope and transmission electron microscope （TEM） ; Semiquantitative reverse transcription - PCR （ RT - PCR） techniques were used to detect the HLA - E mRNA in cytotrophoblasts. Results： Cytotrophoblasts shown round, single core. Microvil lus and cytoplasm were abundant, endocytoplasmic reticulum dilate obviously, glycogenosome, marrow corpuscle and lipid droplet were affluent. HLA - E was expressed on the isolated cytotrophoblast. Conclusion： Highly purified cytotrophoblasts are obtained from term human placenta digested with trypsin, collagenase Ⅰ and density gradient separation percoll, and observed ultrastructure with TEM. Similarly HLA -E is expressed on the cytotrophoblasts, which may be important factor in the maternal- fetal interface.