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目的:建立亚洲人早孕绒毛滋养细胞株以及其相关生物学特征的分析研究。方法:采用胰酶和胶原酶I混合消化法消化绒毛组织,通过光镜、扫描电镜、透射电镜、免疫组化和免疫荧光激光共聚焦的方法对细胞的形态、超微结构、蛋白骨架和功能指标进行特征研究。结果:大多数原代细胞在体外培养至5~7代,产生自然凋亡的现象;少数细胞生长成为永生细胞。细胞检测中角蛋白18、波形蛋白、角蛋白7、移动相关蛋白、表皮生长因子受体、滋养细胞趋化因子和胎盘碱性磷酸酶呈阳性,而人绒毛膜促性腺激素和胎盘生乳素为阴性。结论:采用胰酶和胶原酶I混合消化法可获得较纯的稳定的滋养细胞并可确定该细胞株为具有部分内分泌功能的绒毛滋养细胞株,为进一步开展宫内传播性疾病的实验研究,建立稳定的细胞模型。
OBJECTIVE: To establish an analysis of the trophoblastic trophoblast cells in early pregnancy in Asia and its related biological characteristics. Methods: Villus tissues were digested by trypsin and collagenase I digestion. The morphology, ultrastructure, protein skeleton and function of cells were observed by light microscope, scanning electron microscope, transmission electron microscope, immunohistochemistry and immunofluorescence confocal laser scanning microscope Indicators for characterization. Results: Most of the primary cells were cultured in vitro to 5-7 generations, resulting in the phenomenon of natural apoptosis; a small number of cells grow into immortalized cells. Cytokeratin 18, vimentin, keratin 7, movement related proteins, epidermal growth factor receptor, trophoblast chemotactic factor, and placental alkaline phosphatase were positive for cell detection while human chorionic gonadotropin and placental lactogen were negative. CONCLUSION: Pure and stable trophoblast cells can be obtained by mixed digestion with trypsin and collagenase I, and the trophoblast cells can be confirmed as partial trophoblast cells with partial endocrine function. In order to further study the intrauterine transmission disease, Establish a stable cell model.