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目的:建立一种经济、高效的人足月胎盘滋养细胞分离培养方法。方法:收取正常足月剖宫产患者胎盘组织,DNA酶联合胰酶反复消化、网筛过滤、不连续密度梯度分离液分离纯化细胞;免疫细胞化学法鉴定细胞来源;对所培养的滋养细胞进行形态学观察;CCK-8增殖实验分析细胞体外增殖活力。结果:利用此法可成功体外培养胎盘滋养细胞,一次操作可获得大于107个细胞。体外培养时,滋养细胞增殖能力低下,3h开始贴壁,24h后逐渐开始融合成合体滋养细胞。结论:本研究改进了既往足月胎盘滋养细胞原代培养的方法,简化操作步骤,缩短操作时间,并且一次性可获得大量且纯度较好的胎盘滋养细胞。
Objective: To establish an economical and efficient method of human placental trophoblast isolation and culture. Methods: The placental tissues of normal full-term cesarean section patients were collected, digested with DNase and trypsin, and sieved through a mesh filter. The cells were separated and purified by discontinuous density gradient. The cell sources were identified by immunocytochemistry. The cultured trophoblasts Morphological observation; CCK-8 proliferation assay of cell proliferation in vitro activity. Results: Using this method, we can successfully culture placental trophoblast cells in vitro and obtain more than 107 cells in a single operation. In vitro culture, trophoblastic proliferation was low, 3h began adherent, gradually began to integrate into trophoblast 24h. CONCLUSION: This study improves previous methods of primary term placental trophoblast primary culture, simplifies the procedure, shortens the operation time, and can obtain a large number of placental trophoblast cells with good purity at one time.