摘要
目的 探讨小鼠胚胎肝组织中的 ,干细胞抗原 1阳性的细胞 (Sca 1+ 细胞 )向肾组织上皮细胞分化的潜能。方法 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 10Gy剂量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 ,取肾脏组织固定 ,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肾脏组织内供体小鼠胎肝Sca 1+细胞的分布和横向分化情况。结果 在肾小管上皮内和肾小球边缘 ,分别发现存在Y染色体阳性的供体来源的细胞 ,同时呈现肾组织的部分特征 ,表型分别为RCA+ 、CYP+ 1A2 、CD4 5 -、F-4/ 80 和波形蛋白阳性、CD-45、F-4/ 80 。结论 小鼠胎肝Sca 1+ 细胞 (其中绝大部分是造血干细胞 ) 。
Objective To explore the plasticity and potential of transdifferentiation of stem cell antigen-1 positive cells (Scan-1 + cells) from murine fetal liver into renal cells. Methods The Scan-1 + cells from the livers of male murine fetuses aged 14.5 days were separated with Magnetic cell sorting (MACS) technology. 2×10 3 Scan-1 + cells from the male murine fetuses of the same litter were transplanted through the caudal vein into 6 female mice of the same line aged 8～12 weeks irradiated with the lethal dosage of γ ray from 60 Co source (10 Gy). 6 mice were transplanted with adult wale mariue bone marrow sca-1 + cells and used as experimental controls. Three mice just undergoing irradiation without transplantation was used as blank controls. Sixty days after the transplantation the mice were killed and their kidneys were taken out. Fluorescence in situ hybridization (FISH) test with Y chromosome probe was used to detect cells originated from the source of male donors. Histochemistry with biotin-labeled ricinus communis agglutinin (RCA) and vimentin and immunohistochemistry with antibody of white blood cell common antigen CD45 and antibody of macrophage F 4/80 were used. Results 3 control mice and 2 of the 6 mice transplanted with fetal liver cells died 12, 14, and 15 days and 20 and 26 days respectively after irradiation. In the mice transplanted with Sca-1 + cells only one died 14 days after irradiation and the remaining mice survived to the date to be sampled. FISH technique showed cells with Y chromosome in the epithelium of renal tubules of the female mice transplanted with Sca-1 + cells. RCA +/CYP + 1A2 /CD - 45 /F - 4/80 and vimentin +/CD - 45 /F - 4/80 cells were found in the epithelial cells of renal tubules and glomeruli respectively. Conclusion Scan-1 + cells most of which are hematopoietic stem cells from murine fetal liver can transdifferentiate into renal cells.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2003年第19期1686-1690,共5页
National Medical Journal of China
