摘要
目的:建立B超引导下胎羊体内人干细胞移植模型,及移植细胞向肝脏细胞体内分化的检测方法。方法:通过B超引导下的穿刺,将(0.5～3)×107人干细胞移植入发育50～60d的胎羊体内(n=6);移植后110～120d取材分析,用人HLADR引物PCR检测人源性细胞的嵌合;用人特异性肝细胞、白蛋白、CK18抗体免疫组化染色及Alu探针原位杂交,检测移植细胞向肝脏细胞的分化。结果:在移植的6只胎羊中,5只顺利生产;PCR检测显示5只实验羊肝脏组织中均有人源性细胞的嵌合;免疫组化及Alu探针原位杂交显示人源性细胞分布于门管区周围和肝小叶中,表达白蛋白、CK18等肝细胞分化标志;Alu探针阳性细胞也分布于胆管上皮中。结论:胎羊体内干细胞移植模型及其向肝脏细胞分化的检测,可以有效地显示人干细胞体内向肝脏细胞分化的潜能。
Objective：To establish an animal model of transplanting human stem cells into fetal sheep under B ultrasound guidance, and to detect the in vivo differentiation of stem cells into liver cells. Methods： Human stem cells（0.5-3） × 10^7 were transplanted into sheep fetuses on 50-60 developing days （n =6） through puncture under B ultrasound guidance and the livers were sampled and analyzed 110 120 days later. Human cells engrafted in the samples were detected by PCR analysis with human specific HI.A-DR primers. In vivo differentiation of human stern cells into liver cells was analyzed by immunohistochemical staining with human-specific antibodies against human hepatocyte, Albumin and CK18, and was confirmed by in situ hybridization （ISH） of human-specific Alu probe. Results： Human stem cells were successfully transplanted into 6 fetal sheep and 5 lambs were born and lived. PCR analysis showed that there was human-derived cell engraftment in the liver of the 5 lambs. Immunohistochemical staining and ISH with Alu probe showed human-derived hepatocytes in the lobes or around periportal area, expressing albumin and CK18. Alu ISH also showed there were positive cells in the epithelia of bile ducts. Conclusion： Transplanting human stem cells into fetal sheep and the analysis of stem cell differentiation is a safe and effective system to display the differentiation potential of human stem cells into liver cells.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2005年第10期1120-1123,共4页
Academic Journal of Second Military Medical University
基金
国家高新技术发展规划("863"计划)课题(2001AA216121)
国家重点基础研究规划("973"计划)课题(2001CB509904)
关键词
干细胞移植
肝细胞
细胞分化
stem cell transplantation
hepatocytes
cell differentiation
