体外培养的人滋养层细胞的鉴定  

Preliminary identification of cultured human trophoblasts

作　　者：李珍[1] 黄晓峰[1] 王春梅[1] 徐德忠[2] 门可[2] 王安辉[2] 杨家骥[1] 于华[1] 陈丹[1] 黄小军[1] 王爽[1] 

机构地区：[1]第四军医大学基础部电子显微镜中心,陕西西安710033 [2]第四军医大学预防医学系流行病学教研室,陕西西安710033

出　　处：《第四军医大学学报》2005年第14期1249-1252,共4页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30230320)

摘　　要：目的:对来源于人绒毛组织,体外分离、培养的细胞进行鉴定.方法:复苏已经过纯化和传代培养的人胎盘滋养层细胞继续培养,采用倒置显微镜观察细胞的大体形态、HE染色观察细胞的核浆情况、扫描电镜观察细胞的表面结构、透射电镜观察细胞特有的超微结构、免疫荧光组织化学结合激光扫描共聚焦观察该细胞的细胞角蛋白18(CK18)和波形蛋白(vimentin,Vim)、人绒毛膜促性腺激素(hCG)和人胎盘催乳素(hPL)的表达.结果:该细胞呈不规则多角形片状、单层铺展生长,表面有丰富的微绒毛,细胞之间形成桥粒连接,细胞质内有丰富的线粒体、糖原颗粒和发达的高尔基体、以及脂滴等上皮来源细胞的形态特征;所有培养细胞都表达细胞角蛋白18、而波形蛋白细胞只在部分细胞表达,人绒毛膜促性腺激素和人胎盘催乳素免疫反应阳性物质在细胞质表达.结论:该细胞具有上皮来源细胞的一般形态特征和细胞骨架,以及滋养层细胞特有的激素分泌功能.初步认为该细胞为人胎盘滋养层细胞.AIM: To identify the characteristics of cultured cells from human villous. METHODS: The purified and subcultured cells obtained from human placentae villous were resuscited and cultured. The cellular morphology was observed by inverted phase contrast microscope and the nucleus and cytoplasm were observed by staining of haematoxylin and eosin (HE). The superficial structure was observed by scanning electron microscope (SEM) and the intraceuular ultrastructures were revealed by transmission electron microscope (TEM). The immunofluorescent histochemistry (IH) combined with laser scanning confocal microscopy (LSCM) was used to examine the expression of cytokeratin 18 (CK18), vimentin (Vim), β-human chorionic gonadotropin (β-hCG) and human placental lactogen (hPL). RESULTS: The cultured cells appeared as patchy spreading and irregular polygonal with big oviform nucleus and transparent cytoplasm. SEM observation showed that abundant microvilli existed on the cell surfaces. The typical desmosomes between cells were observed with TEM. The numerous microvilli were also found on their surfaces. The abundant mitochondria, glycogen, developed Golgi complexes, a few of endoplasmic reticulum and many lipid droplets appeared in cytoplasm. Cytokeratin 18 expressed in all the cells cultured and vimentin expressed only in some of the cells. hCG and hPL immunoreactive positive substances were distributed in cytoplasm. CONCLUSION: The cellular morphological features and cytoskeleton of the cultured cells correspond with those of epithelial cells and they secrete the hormones of trophoblast, indicating the cultured cells in vitro are derived from interstitial trophoblast.

关 键 词：滋养层/细胞学 滋养层/超微结构 显微镜检查 电子 扫描透射 显微镜检查 共焦 

分 类 号：R322.65[医药卫生—人体解剖和组织胚胎学]