人胎盘绒毛膜间充质干细胞分离培养:组织块法的优化

发布时间：2018-06-12 02:13

本文选题：胎盘 + 绒毛膜　；参考：《中国组织工程研究》2017年21期

【摘要】：背景:关于人胎盘绒毛膜间充质干细胞分离培养方法的研究很多,但是如何简便、快捷获得大量原代培养间充质干细胞的问题尚未解决。目的:探讨优化人胎盘绒毛膜间充质干细胞体外分离培养组织块法的最佳方法。方法:无菌条件下,取正常足月剖宫产胎盘绒毛膜,用匀浆器处理组织块,采用组织块法贴壁分离培养人胎盘绒毛膜间充质干细胞,经初次培养的组织块进行再次接种培养。结果与结论:用电动匀浆器处理人胎盘绒毛膜省时省力,组织分散度和去除红细胞效果好。初次培养和再次培养获得细胞的平均时间分别为(17.73±1.14)d和(10.03±1.30)d。每个Φ100 mm培养皿获得的原代细胞分别为(6.97±0.98)×10~5个和(13.82±1.44)×10~5个。获得的贴壁细胞呈典型的成纤维细胞形态,呈平行或漩涡状生长,高表达CD90、CD73、CD105,而CD45、CD34、CD14、CD19、HLA-DR为阴性。细胞可向成骨、成脂方向诱导分化。结果表明优化的方法能够简便、快捷地处理人胎盘绒毛膜组织,获得大量的原代间充质干细胞。
[Abstract]:Background: there are many studies on isolation and culture methods of human placental chorionic mesenchymal stem cells, but the problem of how to obtain a large number of primary cultured mesenchymal stem cells quickly and simply has not been solved. Aim: to study the optimal method for isolation and culture of human placental chorionic mesenchymal stem cells in vitro. Methods: human placental chorionic mesenchymal stem cells were isolated and cultured with tissue block adherent method under aseptic condition. The chorionic chorion of normal full-term caesarean section was treated with homogenizer. The first cultured tissue mass was inoculated again. Results & conclusion: the treatment of human placenta chorion with electric homogenate was time saving, tissue dispersion and erythrocyte removal were better. The average time of primary culture and reculture was 17.73 卤1.14 days and 10.03 卤1.30 days, respectively. The primary cells were 6.97 卤0.98 脳 10 ~ 5 and 13.82 卤1.44 脳 10 ~ 5 per 桅 100mm culture dish, respectively. The adherent cells showed typical fibroblast morphology and grew in parallel or whirlpool shape with high expression of CD90, CD73 and CD105, while CD45, CD34, CD14, CD19 and HLA-DR were negative. Cells can be osteoblast, adipogenic direction induced differentiation. The results showed that the optimized method was simple and rapid in processing human placental chorionic tissues and a large number of primary mesenchymal stem cells were obtained.
【作者单位】：广州医科大学附属广州市妇女儿童医疗中心;
【基金】：国家自然科学基金项目(81102248) 广东省医学科研基金(B2010274) 广东省科技计划项目(2014A020 212027) 广州市妇女儿童医疗中心儿科研究所基金(YIP-2016-014)~~
【分类号】：R329.2

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