小鼠胎肝Sca-1^+细胞分化为骨骼肌细胞

Differentiation of murine fetal liver derived Sca-1^+ cells into skeletal muscle cells in vivo

探讨小鼠胚胎肝组织中干细胞抗原 1阳性的细胞 (Sca 1+ 细胞 )向骨骼肌细胞分化的潜能。取 14 5d的小鼠胎肝 ,制作细胞悬液 ;用单克隆免疫磁珠细胞分离技术分离Sca 1+ 细胞 ;用PCR鉴别Y染色体性别决定区域 (SRY)基因序列 ;将 2× 10 3 个雄性小鼠Sca 1+ 细胞输注给经致死剂量 (10Gy) 60 钴全身照射的雌性小鼠体内 ;于移植后 2个月 ,处死受体小鼠 ,取骨骼肌组织固定、制片 ;用免疫组织化学染色和荧光原位杂交检测雌性受体小鼠骨骼肌组织内供体小鼠胎肝Sca 1+ 细胞向骨骼肌细胞分化情况。结果在骨骼肌组织内发现存在Y染色体阳性的供体来源的细胞 ,同时呈现骨骼肌组织的部分特征 ,表型为Dystrophin+ /Flt 1-/CD4 5 -F4 / 80 -。提示小鼠胎肝Sca 1+ 细胞 (其中绝大部分是造血干细胞 )具有向骨骼肌组织细胞分化的潜能。

To explore the differentiation potential of stem cell antigen-1 positive cells (Sca-1 + cells) from murine fetal liver into skeletal muscle cells in vivo. The Sca-1 + cells from the murine livers of 14.5 days were separated by Magnetic cell sorting (MACS) technology. 2×10 3 of Sca-1 + cells from the male were transplanted through the tail vein into 6 female mice of 8～12 weeks irradiated with the lethally dosage of γ ray from Co 60 source (10Gy) .Two months later,the mice were killed and their skeletal muscles were taken out to make slides. Fluorescence in situ hybridisation (FISH) test with Y chromosome probe was used to detect the sex of cells。Immunohistochemistry with antibodies of dystrophin 、Flt-1 、 white blood cell common antigen CD 45 and macrophage F 4/80 were used to detect tissue characteristics of skeletal muscles. The results showed that the cells with Y chromosome in the skeletal muscle of female mice transplanted with Sca-1 + cells with showed the characteristics of Dystrophin + /Flt-1 - CD -45 F -4/80 at the same time. It is suggested that Sca-1 + cells from murine fetal liver can differentiate into skeletal muscle cells.