背景:脐带间充质干细胞治疗肝衰竭已经取得了一定的疗效,但是肝衰竭患者的血清微环境对脐带间充质干细胞分化的影响尚未见报道.目的:探讨慢加急性肝衰竭患者血浆置换前、后血清对脐带间充质干细胞向功能肝细胞分化的影响.方法:采用组织块贴壁培养法获得脐带间充质干细胞并进行细胞形态及表型鉴定;取第3代脐带间充质干细胞,用含有肝衰竭患者血浆置换前、后血清的 α-MEM培养基诱导培养,常规胎牛血清培养基为对照组,显微镜下观察各组细胞的形态,免疫组化检测细胞甲胎蛋白、白蛋白和细胞角蛋白18的表达水平.结果与结论:①组织块贴壁培养法能够获得大量高纯度的脐带间充质干细胞,细胞高表达CD44,CD73,CD90,CD105,不表达CD45,符合间充质干细胞的表型特征;②慢加急性肝衰竭患者血清可改变脐带间充质干细胞的形态,并诱导脐带间充质干细胞表达甲胎蛋白、白蛋白和细胞角蛋白18.血浆置换后血清组3种蛋白阳性率高于血浆置换前血清组,差异有显著性意义(P<0.05);③结果表明,肝衰竭患者血浆置换后血清更加有利于脐带间充质干细胞生长及向肝细胞分化.%BACKGROUND: Umbilical cord mesenchymal stem cells (UC-MSCs) have made certain curative effect on hepatic failure, but little is reported on the effect of hepatic failure patient's serum microenvironment on UC-MSCs differentiation ability. OBJECTIVE: To investigate the effect of hepatic failure patient's serum on the differentiation of umbilical cord mesenchymal stem cells into hepatic cells. METHODS: UC-MSCs were isolated by tissue adherent method and the cell morphology and phenotype identified by microscope and flow cytometry. Alpha-MEM media with serum before/after plasmapheresis were used to culture the hirdgeneration of UC-MSCs, and regular fetal bovine serum culture medium acted as control group. Inverted microscope was used to observe the cell morphology in three groups. Immunohistochemical method was used to measure expression level of alpha fetoprotein, albumin and cytokeratin 18. RESULTS AND CONCLUSION: A great amount of high-purity UC-MSCs could be obtained using tissue adherent method, which highly expressed CD44, CD73, CD90, CD10, but did not express CD45. Hepatic failure patient's serum could change the morphology of UC-MSCs and induce UC-MSCs to express alpha fetoprotein, albumin and cytokeratin 18. The positive expression of alpha fetoprotein, albumin and cytokeratin 18 was significantly increased after plasmapheresis (P < 0.05). To conclude, hepatic failure patient's serum after plasmapheresis exert more benefits to induce UC-MSCs to differentiate into hepatic cells than that before plasmapheresis.
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