| 改良的高纯度人早孕绒毛膜滋养层细胞培养方法 |
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| 引用本文： | 陈莉娟,周浩,朱剑文,邹丽.改良的高纯度人早孕绒毛膜滋养层细胞培养方法[J].中国组织化学与细胞化学杂志,2008,17(3). |
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| 作者姓名： | 陈莉娟 周浩 朱剑文 邹丽 |
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| 作者单位： | 1. 华中科技大学同济医学院附属协和医院妇产科,武汉,430022
2. 华中科技大学同济医学院附属协和医院血液病研究所,武汉,430022 |
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| 摘 要： | 目的培养纯度较高的人早孕绒毛膜滋养层细胞,为研究胎盘绒毛在妊娠期间的作用及其机制提供细胞学基础。方法胰蛋白酶-DNA酶消化法分离培养绒毛膜滋养层细胞,再运用流式细胞仪细胞分选获得高表达HLA-G的人早孕绒毛膜滋养层细胞。流式细胞术检测分选前后原代培养细胞HLA-G的表达率,相差显微镜观察滋养层细胞形态学特点,免疫荧光显微镜鉴定细胞来源,台盼蓝染色检测细胞活力。结果通过流式细胞检测,分选前的原代培养细胞体系中HLA-G的表达率为86.5%,经过分选带有PE荧光/HLA-G阳性表达的原代培养细胞后,其纯度可达98.0%。倒置相差显微镜下,可见细胞为上皮样细胞形态,呈片状铺展生长。细胞角蛋白染色阳性,波形蛋白染色阴性,表明细胞性质为上皮来源的绒毛膜滋养层细胞。台盼兰排斥试验检测细胞活力,细胞活力良好,存活率超过92%。结论该方法可以有效获得高纯度的,具有生物学活性的人早孕绒毛膜层细胞,为在体外研究生理妊娠及病理妊娠中滋养细胞的作用提供了一种改良的技术手段。
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| 关 键 词： | 滋养层细胞 HLA-G 细胞培养 |
AN IMPROVED METHOD OF PRIMARY CULTURE FOR PURIFIED HUMAN FIRST TRIMESTER PREGNANCY TROPHOBLAST CELLS |
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| Chen Lijuan,Zhou Hao,Zhu Jianwen,Zou Li.AN IMPROVED METHOD OF PRIMARY CULTURE FOR PURIFIED HUMAN FIRST TRIMESTER PREGNANCY TROPHOBLAST CELLS[J].Chinese Journal of Histochemistry and Cytochemistry,2008,17(3). |
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| Authors: | Chen Lijuan Zhou Hao Zhu Jianwen Zou Li |
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| Institution: | Chen Lijuan1,Zhou Hao2,Zhu Jianwen1,Zou Li1(1Department of Obsterics , Gynecology,Union Hospital,Tongji Medical College,Huazhong University of Science , Technology,Wuhan 430030,2Institute of Hematology,Wuhan 430022,China) |
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| Abstract: | Objective To explore an improved method to prepare highly purified human trophoblast cells from human placenta at the first trimester of normal pregnancy for further study on the function of human placenta villi.Methods The standard trypsin-DNase dispersion method was used to isolate the human first trimester trophoblast cells.Then the cultured cells were stained with PE conjugated antibody and applied to flow cytometry for cell sorting.The sorted HLA-G positive cells were cultured and their purity was dete... |
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| Keywords: | Trophoblast cell HLA-G Cell culture |
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