成人骨髓干细胞向肝细胞诱导分化的实验研究

发布时间：2019-01-09 18:17

【摘要】：研究背景和目的：对于各种终末期肝病，原位肝移植(orthotopic liver transplantation，OLT)理论上应是最为理想的治疗选择，但有一定的局限性。肝细胞移植与原位肝移植比较，具有操作简单、可重复性强、一肝可以多用、供肝细胞免疫原性低且可冻存、移植失败或产生免疫排斥后对受体影响小等优点；此外，肝细胞在体外还可以进行基因修饰，是一个良好的基因治疗载体。近年来，肝细胞移植在理论与技术方面均取得了很大进展，在许多方面都呈现出成为OLT替代治疗方案的潜力。人骨髓来源的造血干细胞(hemtopoietic stem cell，HSCs)可以分化为骨、软骨、脂肪和肌细胞等多种细胞。目前研究发现，在适当的条件下，骨髓干细胞也可以分化为具有肝细胞功能的肝细胞样细胞。目前，，关于成人骨髓干细胞向肝细胞诱导分化的国际报道有一篇，国内尚无相关报道；关于诱导分化细胞的冻存方法国内外尚无相关报道。本实验主要研究目的：1)在体外诱导人骨髓来源的干细胞向肝细胞分化；2)对诱导分化的肝细胞进行形态和功能鉴定；3)观察经诱导分化的肝细胞冻存、复苏后的功能变化。实验方法： 1．采用体外细胞培养的方法培养人的骨髓干细胞，在含有肝细胞生长因子(hepatocyte growth factor，HGF)、白细胞抑制因子(Leukocyte inhibitory factor，LIF)、纤维母细胞生长因子(Gibroblast growth factor，FGF)等的诱导培养基中进行细胞
[Abstract]:Background and objective: orthotopic liver transplantation (orthotopic liver transplantation,OLT) should be the most ideal treatment for various end-stage liver diseases in theory, but it has some limitations. Compared with orthotopic liver transplantation, hepatocyte transplantation has the advantages of simple operation, strong reproducibility, low immunogenicity of donor liver cells, low immunogenicity, little influence on recipients after transplantation failure or immune rejection. In addition, hepatocytes can also be genetically modified in vitro, which is a good gene therapy vector. In recent years, hepatocyte transplantation has made great progress in both theory and technology, and it has the potential to become an alternative therapy for OLT in many aspects. Human bone marrow derived hematopoietic stem cells (hemtopoietic stem cell,HSCs) can be differentiated into bone, cartilage, fat and muscle cells. It has been found that bone marrow stem cells can also differentiate into hepatocyte-like cells under suitable conditions. At present, there is an international report on the induction and differentiation of adult bone marrow stem cells into hepatocytes, but there is no related report at home and abroad, and there is no relevant report at home and abroad on the cryopreservation methods of inducing differentiated cells. The main objectives of this study were: 1) to induce the differentiation of human bone marrow derived stem cells into hepatocytes in vitro, 2) to identify the morphology and function of the differentiated hepatocytes. 3) to observe the functional changes of differentiated hepatocytes after cryopreservation and resuscitation. Methods: 1. Human bone marrow stem cells were cultured in vitro with hepatocyte growth factor (hepatocyte growth factor,HGF) and leukocyte inhibitory factor (Leukocyte inhibitory factor,LIF). Fibroblast growth factor (Gibroblast growth factor,FGF et al.
【学位授予单位】：中国协和医科大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R329.2

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