【摘要】：研究背景和目的： 對(duì)于各種終末期肝病，原位肝移植(orthotopic liver transplantation，OLT)理論上應(yīng)是最為理想的治療選擇，但有一定的局限性。肝細(xì)胞移植與原位肝移植比較，具有操作簡(jiǎn)單、可重復(fù)性強(qiáng)、一肝可以多用、供肝細(xì)胞免疫原性低且可凍存、移植失敗或產(chǎn)生免疫排斥后對(duì)受體影響小等優(yōu)點(diǎn)；此外，肝細(xì)胞在體外還可以進(jìn)行基因修飾，是一個(gè)良好的基因治療載體。近年來(lái)，肝細(xì)胞移植在理論與技術(shù)方面均取得了很大進(jìn)展，在許多方面都呈現(xiàn)出成為OLT替代治療方案的潛力。 人骨髓來(lái)源的造血干細(xì)胞(hemtopoietic stem cell，HSCs)可以分化為骨、軟骨、脂肪和肌細(xì)胞等多種細(xì)胞。目前研究發(fā)現(xiàn)，在適當(dāng)?shù)臈l件下，骨髓干細(xì)胞也可以分化為具有肝細(xì)胞功能的肝細(xì)胞樣細(xì)胞。目前，，關(guān)于成人骨髓干細(xì)胞向肝細(xì)胞誘導(dǎo)分化的國(guó)際報(bào)道有一篇，國(guó)內(nèi)尚無(wú)相關(guān)報(bào)道；關(guān)于誘導(dǎo)分化細(xì)胞的凍存方法國(guó)內(nèi)外尚無(wú)相關(guān)報(bào)道。 本實(shí)驗(yàn)主要研究目的：1)在體外誘導(dǎo)人骨髓來(lái)源的干細(xì)胞向肝細(xì)胞分化；2)對(duì)誘導(dǎo)分化的肝細(xì)胞進(jìn)行形態(tài)和功能鑒定；3)觀察經(jīng)誘導(dǎo)分化的肝細(xì)胞凍存、復(fù)蘇后的功能變化。 實(shí)驗(yàn)方法： 1．采用體外細(xì)胞培養(yǎng)的方法培養(yǎng)人的骨髓干細(xì)胞，在含有肝細(xì)胞生長(zhǎng)因子(hepatocyte growth factor，HGF)、白細(xì)胞抑制因子(Leukocyte inhibitory factor，LIF)、纖維母細(xì)胞生長(zhǎng)因子(Gibroblast growth factor，F(xiàn)GF)等的誘導(dǎo)培養(yǎng)基中進(jìn)行細(xì)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.
【學(xué)位授予單位】：中國(guó)協(xié)和醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2005
【分類(lèi)號(hào)】：R329.2

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