摘要
目的研究分离胎儿骨髓源性亚全能干细胞及体内、外向肝细胞分化的潜能。方法利用密度梯度离心结合免疫磁珠方法分离胎儿骨髓源性亚全能干细胞,体外培养鉴定,诱导分化。制备肝功能衰竭重度联合免疫缺陷小鼠(severecombinedimmunodeficiency,SCID)模型。肝原位输注106左右CD105+细胞,对照组分别输注106左右的CD105-细胞或同等体积的培养液。移植细胞后2d、7d、1个月、3个月时分别取3只鼠的血清及组织标本行肝功能和病理学、免疫组织化学检测。结果免疫磁珠筛选后的免疫细胞化学检测CD105呈弱阳性表达;细胞在对数生长期的倍增时间为30h左右;约传10代后进入衰退期;SCID鼠移植细胞3个月后用鼠抗人白蛋白抗体检测小鼠肝脏中的人白蛋白,可见有点状或小灶状表达。结论来源于胎儿骨髓的亚全能干细胞可以在体外及肝脏微环境下转化为肝细胞样细胞。因此进一步深入研究组织微环境在细胞转化中的机制有十分重要的意义,将为开发诱导胎儿骨髓亚全能干细胞的多向分化潜能提供理论依据。
Objective To study the isolation of human fetal bone marrow postembryonic pluripotent stem cells and their differentiation to hepatocytes in vivo and in vitro. Methods The human fetal bone marrow postebryonic pluripotent stem cells were isolated by density gradient centrifugation and immune micromagnetic beads technique, then identified the culture cells and induced to defferentiate in vitro. The hepatic failure model was established with severe combined immunodeficiency （SCID） mice. In experimental group 106 CD 105^＋ cells were infused into mice liver, and 106 CD105^- cells or same volume culture medium were infused into mice liver in control group. Serum and tissue sample of three mice serume were taken at 2 days,7 days, 1 month and 3 months respectively after cell transplantation for examination of liver function ,pathology and immunocytochemical assay. Results The cells screened by immunomagnetic beads expressed weak positive of CD105 in immunocytechemical assay. The doubling time of the cells in logarithmic growth period was around 30 hours, the cells entered decline period after expanding for 10 generations. Dotted or small focal expressions were found in SCID mouse liver when detecting the human serum albumin with mouse anti-human albumin antibody after transplantation of the stem cells in 3 months. Conclusion The human fetal bone marrow postembryonic pluripotent stem cells can differentiate to hepatocyte-like cells in vitro or in hepatic microcirculation. Thus further study on the mechanism of cell transformation in tissue microcirculation is a topic of important significance,it will provides the theoretical basis to develop the induction of multi-directed differentiation in human fetal bone marrow postembryonic pluripotent stem cells.
出处
《生物医学工程与临床》
CAS
2007年第1期63-65,F0003,共4页
Biomedical Engineering and Clinical Medicine
基金
国家高新技术研究发展计划"(863"计划)(2001AA216051)
关键词
骨髓
亚全能干细胞
肝细胞样细胞
胎儿
bone marrow
postembryonic pluripotent stem cell
hepatocyte-like cell
human fetal
