Preliminary identification of cultured human trophoblasts

    LI Zhen, HUANG Xiaoª²Feng, WANG Chunª²Mei, XU Deª²Zhong, MEN Ke, WANG Anª²Hui, YANG Jiaª²Ji, YU Hua, CHEN Dan, HUANG Xiaoª²Jun, WANG Shuang

    1Electron Microscopic Center, Fourth Military Medical University, Xi¬ðan 710033, 2Department of Epidemiology, Faculty of Preventive Medicine, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To determine the characteristics of human trophoblasts in vitro. METHODS£ºThe purified and subcultured cells which come from human placentae villous were resuscited and cultured.The general form of cells were observed by interestinverted phase contrast microscope , the nucleoplasm of cells were observed by taining of haematoxylin and eosin (HE). The superficial structure of cells were observed by scanning electron microscope (SEM), The ultrastructure of cells was revealed by transmission electron microscope (TEM), the technique of Immunofluorescent histochemistry(IH) was used to examine the expression of cytokeratin 18 (CK18) and vimentin (Vim) of cellular skeleton,¦Âª²human Chorionic Gonadotropin (¦Âª²hCG) and human Placental Lactogen (hPL) with laser scanning confocal microscopy (LSCM). RESULTS: Light microscopic examination revealed patchy spreading and irregular polygonal cells with big and oviform nuclear and transparent cytoplasm. SEM observation showed that abundant microvilli on the cell surfaces, cells were patchy spreading and irregular polygonal, gathered into pieces and maybe changed into syncytiotrophoblasts. Observed by TEM, cells showed typical structural characteristics of trophoblasts, such as the numerous microvilli on cell surface, desmosome between cells, abundant mitochondria, glycogen, developed Golgi complex,a few of endoplasmic reticulum and many lipid droplets in cytoplasm. Observed by LSCM, cytokeratin 18 was positive, vimentin was positive in part of cells, hCG expression was positive and hPL expression was positive and distributed in cytoplasm. CONCLUSION: The cells cultured in vitro maybe derived from interstitial trophoblast. They are patchy spreading and irregular polygonal, which correspond with the morphological characteristics of epithelial cells cultured in vitro. These characteristics of ultrustructure and the expressings of characteristic antibodies are distinguished with others and confirm that it can synthesize and secrete some substances. ......