人脐带间充质干细胞的分离技术优化及其快速检测体系的建立

Optimization of human umbilical cord mesenchymal stem cell isolation and establishment of a rapid detection system

目的研究人脐带间充质干细胞(UCMSC)的分离技术优化及其快速检测体系的建立效果。方法选取2018年3月-2018年6月,于我院进行分娩的20例健康新生儿脐带组织标本作为研究对象,将其以随机抽签法等分成2份。其中1份采用传统的贴壁法进行细胞的分离培养,并采用传统细胞检测方式进行人UCMSC的检测,记为对照组。另外1份则采用优化分离技术进行细胞的分离培养,并采用快速检测体系对人UCMSC进行检测,记为研究组。比较两组细胞生长情况以及得率情况,第3代、6代、9代细胞的增殖情况,细胞检测时间。结果研究组初次见贴壁细胞时间、80%贴壁/可传代时间相比对照组较低,而细胞得率相比对照组较高(均P<0.05)。研究组第3代、6代、9代细胞4d、8d时的细胞数均高于对照组(均P<0.05)。研究组细胞检测时间为0.81±0.15d,相比对照组的2.34±0.21d较低,差异明显(P<0.05)。结论优化法应用于人UCMSC的分离培养中效果显著,有利于提高人UCMSC的质量。同时,快速检测体系的建立可显著缩短人UCMSC的检测时间。

Objective To optimize the separation technology of human umbilical cord mesenchymal stem cells(UCMSC)and to analyze the effect of establishing a rapid detection system.Methods Twenty healthy neonatal umbilical cord tissue samples delivered in our hospital from March 2018 to June 2018 were selected as research objects and divided into two parts by random lottery.One of them was isolated and cultured by the traditional adherent method,and the human UCMSC was detected by the traditional cell detection method,which was recorded as the control group.The other one was isolated and cultured using optimized separation technology,and the human UCMSC was detected by rapid detection system,which was recorded as the study group.The growth and yield of cells in the two groups,the proliferation of cells in the third,sixth and ninth generation,and the detection time of cells were compared.Results Compared with the control group,the first time of adherent cells and 80%adherent/passable time in the study group were lower,while the cell yield was higher(all P<0.05).The number of cells in the third,sixth and ninth generation was higher at 4d and 8d than that in the control group(all P<0.05).The cell detection time in the study group was(0.8±0.15)d,which was lower than the control group(2.34±0.21)d,with significant difference(P<0.05).Conclusion The optimization method is effective in the isolation and culture of human UCMSC,which is beneficial to improve the quality of human UCMSC.Meanwhile,the establishment of rapid detection system can significantly shorten the detection time of human UCMSC.