An efficient method of sorting liver stem cells by using immuno-magnetic microbeads

AIM： To develop a method to isolate liver stem cells fast and efficiently. METHODS： Fetal mouse liver cells were characterized by cell surface antigens （c-Kit and CD45/TER119） using flow cytometry. The candidate liver stem cells were sorted by using immuno-magnetic microbeads and identified by -lone-forming culture, RT-PCR and immunofluorescence says. RESULTS： The c-Kit-（CD45/TER119）- cell population with 97.9% of purity were purified by immuno-magnetic microbeads at one time. The yield of this separation was about 6% of the total sorting cells and the cell viability was above 98%. When cultured in vitro these cells had high clone-forming and self-renewing ability and expressed markers of hepatocytes and bile duct cells. Functionally mature hepatocytes were observed after 21 d of culture. CONCLUSION： This method offers an excellent tool for the enrichment of liver stem cells with high purity and viability, which could be used for further studies. It is fast, efficient, simple and not expensive.

瞄准：开发一个方法快并且高效地孤立肝干细胞。方法：胎儿的老鼠肝细胞被房间表面抗原(c 工具包和 CD45/TER119 ) 用流动血细胞计数描绘。候选人肝干细胞被使用免疫磁性的微祷告排序并且由形成克隆的文化， RT-PCR 和免疫荧光试金识别了。结果：c-Kit-(CD45/TER119 )- 有 97.9% 纯净的房间人口同时被免疫磁性的微祷告净化。这分离的收益是排序房间的大约 6% 总数，房间生存能力超过 98% 。什么时候这些房间有的有教养的在试管内高形成克隆、自我更新的能力和 hepatocytes 和胆汁管房间的表示标记。机能上地成熟的 hepatocytes 在文化的 21 d 以后被观察。结论：这个方法与高纯净和生存能力为肝干细胞的丰富提供一个优秀工具，它能被用于进一步的研究。它快、有效、简单、不昂贵。