目的:鉴定小鼠骨髓间充质干细胞(MSCs)的多分化潜能的起因。方法:从成年小鼠骨髓股骨、胫骨中分离MSCs,并进行原代培养,分别于0、2、4、6、8、10 d提取总RNA,通过RT-PCR检测多潜能特征基因和相关因子、3个胚层特征基因的mRNA的表达情况,以判断MSCs的特性。结果:小鼠MSCs中表达多能性标记基因Oct-4和nanog,并且表达与多能性相关的转录因子Klf4和c-Myc。外胚层的nestin、中胚层的SM22a和内胚层的CYP51标记基因均有表达。结论:贴壁纯化的小鼠MSCs除了具有多能性的干细胞,还可能含有各胚层的原始细胞。
Aim:To identify the cause of multi-differentiation potential in mouse bone marrow mesenchymal stem cells.Methods: MSCs were isolated from the femurs and tibiae of the adult mouse and cultured in vitro.Total RNA were extracted from the mouse bone marrow(Day 0) and cultured mouse MSCs at 2,4,6,8 and10 days.The pluripotency marker genes Oct-4 and nanog,pluripotency related transcription factors Klf4 and c-Myc and the marker genes of three embryonal layers,such as CYP51,SM22α and nestin,which are typical genes specific for endoderm,mesoderm and ectoderm respectively,were detected by RT-PCR in order to determine the characteristics of MSCs.Results: The results of RT-PCR showed that mouse MSCs can express mRNA of Oct-4,nanog,Klf4 and c-Myc.Furthermore,in addition to SM22a,RT-PCR revealed the presence of mRNA for CYP51 and nestin during the primary culture of mouse MSCs.Conclusion:Mouse MSCs,which were isolated by adhesion purification,may be consist of pluripotent stern cells and primary cells from the three germ layers.