人胎盘底蜕膜间充质干细胞库的初步构建  

Incipient Establishment of Human Placenta Decidua Basalis-Derived Mesenchymal Stem Cells Bank

作　　者：肖桂清 段训威 唐燕程 郑梦盈 苏爱盟 耿晓波 刁勇[3] XIAO Guiqing;DUAN Xunwei;TANG Yancheng;ZHENG Mengying;SU Aimeng;GENG Xiaobo;DIAO Yong(Fujian Yinfeng Stem Cell Engineering Co.,Ltd,Fuzhou 350100,China;Quanzhou Normal University,Quanzhou 362000,China;School of Medicine,Huaqiao University,Quanzhou 362021,China)

机构地区：[1]福建省银丰干细胞工程有限公司,福州350100 [2]泉州师范学院,泉州362000 [3]华侨大学医学院,泉州362021

出　　处：《中国细胞生物学学报》2021年第6期1257-1264,共8页Chinese Journal of Cell Biology

基　　金：福建省自然科学基金(批准号:2019J05094);福建省科技计划高校产学合作项目(批准号:2018Y4009);福建省中青年教师教育科研项目(批准号:JAT200544)资助的课题。

摘　　要：该文探讨了建立生物学特性稳定的人胎盘底蜕膜间充质干细胞(DB-MSCs)库的可行性,旨在为组织工程种子细胞提供更多来源。该研究采用组织块贴壁法从10例足月人胎盘底蜕膜组织中获取间充质干细胞,并采用STR检测细胞是否均来源于母体组织。对分离得到的DB-MSCs采用–196 ℃低温冻存,并在一定时间复苏培养,用倒置显微镜观察细胞形态,CCK8法检测细胞增殖能力,流式细胞仪分析细胞周期和细胞表面标志物,特定培养基诱导其成脂、成骨、成软骨分化。结果显示,从底蜕膜组织中分离培养扩增获得数目稳定的DB-MSCs。STR分析证明,所得细胞均来源于母体组织,经冻存并复苏后的DB-MSCs形态和表面标志物保持不变,并可稳定扩增8代以上,倍增时间为(2.04±0.25)天,大多数的细胞处于静止期(G_(0)/G_(1)),复苏后的细胞保持了较强的成脂、成骨和成软骨分化能力,在细胞中未发现细菌、真菌、支原体和内毒素等的污染。该研究初步建立了DB-MSCs库,进一步明确了库存细胞的制备流程及其质量评价体系,可为再生医学修复重建组织工程提供较好的种子细胞。The aim of this study was to explore the possibility of establishing the human placenta DB-MSCs (decidua basalis-derived mesenchymal stem cells) bank as to provide an alternative source for the seed cells of tissue engineering.DB-MSCs were harvested from the decidua basalis of 10 cases of full-term human placental by tissue explants adherent method,and STR (short tandem repeat) test was used to identify the cells derived from the maternal placenta surface.The proliferative ability of the cells was detected by CCK8 assay.Flow cytometry was applied to analyze cell cycle and stem cell markers.The results of STR analysis showed that the cells were derived from the maternal placenta.The phenotype and expansion possibility of DB-MSCs after cryopreservation were remained.It could expand for 8 generations.The doubling time was (2.04±0.25) days.Cell cycle analysis showed majority cells stayed in the resting stage (G_(0)/G_(1)).The DB-MSCs kept stronger ability of adipogenesis,osteoblasts and chondrogenic differentiation.The bank of DB-MSCs has been incipiently established.This article further defines the inventory cell preparation process and its quality evaluation system which can provide eligible seed cells for tissue engineering.

关 键 词：底蜕膜 间充质干细胞 生物学特性 多向分化 细胞库 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]