骨髓c-Kit+Lin-细胞移植改善肝纤维化的实验研究
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陶强, Email: taoqiang_2008@126.com

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江西省卫生厅科技计划资助项目(20113098);江西省科技支撑计划资助项目(2010BSA11200)。


Bone marrow c-Kit+Lin? cells transplantation for hepatic fibrosis: an experimental study
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    摘要:

    目的:探讨骨髓c-Kit+Lin-(CD117)细胞在体外扩增后移植治疗小鼠肝纤维化的效果。 方法:采用免疫磁珠分选法分离小鼠骨髓CD117细胞,将细胞在添加了肝细胞生长因子(HGF),干细胞因子(SCF),FLt-3配基(FL),促血小板生成素(TPO),白血病抑制因子(LIF)的无血清培养基中培养扩增;制备同种小鼠的酒精性肝纤维化模型,成模后的小鼠分别尾静脉注射体外扩增的CD117细胞(移植组)或等体积缓冲液(模型组),同时以正常小鼠为对照(正常组),30 d后观察各组小鼠的肝脏病理改变以及肝功能与肝纤维化指标。 结果:添加上述细胞因子的无血清培养基能有效扩增CD117细胞;正常组小鼠肝脏无病理改变,模型组呈明显的肝细胞变性和纤维化改变,移植组较模型组的病变程度明显减轻;与正常组比较,模型组和移植组均有不同程度的血清谷丙转氨酶(ALT),谷草转氨酶(AST),透明质酸(HA),层黏连蛋白(LN)升高和清蛋白(ALB)降低,但移植组上述指标改变较模型组明显减轻,差异均有统计学意义(均P<0.05)。 结论:合适剂量的HGF+SCF+FL+TPO+LIF细胞因子组合能有效扩增CD117细胞;CD117细胞移植对小鼠肝纤维化有治疗作用。

    Abstract:

    Objective: To investigate the therapeutic effect of in vitro amplified bone marrow c-Kit+Lin? (CD117) cells transplantation on hepatic fibrosis in mice. Methods: The bone marrow c-kit+Lin? cells of mice were isolated and purified by using magnetic-activated cell sorting (MACS), and then the cells were cultured and expanded in the serum-free medium supplemented with hepatocyte growth factor (HGF), stem cell factor (SCF), FLt-3 ligand (FL), thrombopoietin (TPO) and leukemia inhibitory factor (LIF). The models of alcohol induced hepatic fibrosis were created in mice of the same strain, and the model mice were treated with CD117 cells (transplantation group) or buffer solution of the same volume (model group) by tail vein injection respectively, and the normal mice were used as control (normal group). Thirty days later, the pathological changes of liver and the parameters concerning liver function and hepatic fibrosis of each group were assessed. Results: The CD117 cells were effectively expanded in the serum-free medium supplemented with the above mentioned factors. No pathological alteration was noted in the liver of normal mice, while the liver of model group presented remarkable hepatocyte degeneration and fibrotic changes, and these alterations were obviously alleviated in transplantation group compared with model group. In both model and transplantation groups, the serum levels of alanine transaminase (ALT), aspartate transaminase (AST), hyaluronic acid (HA) and laminin (LN) increased, and albumin (ALB) decreased with varying degrees compared with normal group, but the alterations of the above parameters in transplantation group were all milder than those in model group, and all differences had statistical significances (all P<0.05). Conclusion: CD117 cells can be effectively expanded in vitro by the proper combination of cytokins (HGF, SCF, FL, TPO and LIF). CD117 cells transplantation has therapeutic effect on hepatic fibrosis in mice.

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张守华|肖菊花|邓庆强|徐维|胡瞒鑫|谌卫龙|陶强.骨髓c-Kit+Lin-细胞移植改善肝纤维化的实验研究[J].中国普通外科杂志,2012,21(7):821-824.
DOI:10.7659/j. issn.1005-6947.2012.07.010

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  • 收稿日期:2011-08-23
  • 最后修改日期:2012-05-03
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  • 在线发布日期: 2012-07-15