中文摘要：

背景：通过动员自身或移植外来的骨髓来源肝干细胞可促进肝再生,但是,在大规模临床应用前,其安全性需要进一步研究。目的：采用含体积分数5%淤胆血清的培养基诱导大鼠骨髓间充质干细胞向肝干细胞方向分化,将这些骨髓来源肝干细胞接种到裸鼠体内,观察其是否具有成瘤性。方法：用含体积分数5%淤胆血清的培养基培养大鼠骨髓间充质干细胞;以免疫荧光法检测白蛋白、甲胎蛋白及细胞角皮素18在诱导后细胞的表达;以糖原染色及尿素合成检测细胞功能。将培养14d的大鼠骨髓来源肝干细胞接种于裸鼠皮下,观察局部有无新生物形成。结果与结论：用含体积分数5%淤胆血清的培养基培养大鼠骨髓间充质干细胞,4d后出现细胞集落,细胞为圆形;7d后集落变大,其周围开始出现多角形细胞;培养14d后可见细胞呈多角形和排列成铺路石样,免疫荧光染色发现这些细胞表达角皮素18、甲胎蛋白和白蛋白,糖原染色显示细胞内有糖原颗粒;培养第12～15天的培养液中尿素氮浓度逐渐升高。经诱导的大鼠骨髓来源的肝干细胞接种到裸鼠皮下,30d后局部未见新生物形成,组织结构未见异常。结果提示用体积分数5%淤胆血清培养基诱导的大鼠骨髓源性肝干细胞可能无成瘤性。

英文摘要：

BACKGROUND：Mobilizing autologous or extraneous bone marrow-derived liver stem cells may promote liver regeneration,however,its safety before the large scale clinical application needs further evaluation.OBJECTIVE：Bone marrow-derived liver stem cells （BDLSCs） were induced by culturing the rat bone marrow mesenchymal cells in the medium containing 5% cholestatic sera,and then were implanted into nude mice to observe the tumorigenicity.METHODS：Rat bone marrow mesenchymal cells （BMSCs） were isolated and incubated in the medium containing 5% cholestatic sera.Immunofluorescent stain was used to detect the expression of albumin,alpha-fetoprotein and cytokeratin18 by the cultured cells.Glycogen and urea synthesis by these cells were analyzed,respectively.BDLSCs following 14 days of culture were incubated in the skin of nude mice to observe neoplasia in local site.RESULTS AND CONCLUSION：Rat BMSCs survived in the medium containing 5% cholestatic serum and formed into small colonies on the fourth day after culture.Seven days later,the colonies expanded and there appeared some polygonal cells in the peripheral area.About 14 days later,these polygonal cells were confluent and presented the shape of cobblestone.Immunofluorescent stain showed that these cells expressed cytokeratin18,albumin and alpha-fetoprotein.Staining for glycogen displayed that glycogen granules were seen in cells.From 12 to15 days after culture,urea nitrogen concentrations in the medium were gradually increased.Rat BDLSCs were incubated in the skin of nude mice.Thirty days later,no neoplasia was found in the local site,and the tissue structure was normal.This result indicated that rat BDLSCs induced with the medium containing 5% cholestatic serum might have not tumorigenicity.