人骨髓间充质干细胞向造血细胞分化潜能的研究

发布时间：2019-02-18 15:24

【摘要】：骨髓间充质干细胞(MSC)是一种具有多向分化潜能的干细胞,在体外培养扩增且能长期维持其遗传性状稳定。造血干细胞移植(HSCT)作为一种重要的生物或细胞治疗,尽管已取得了很好的临床疗效,但供源不足、不能在体外大量扩增等方面原因使得HSCT 的应用仍受多方面的限制。那么,人骨髓MSC(hMSC)能否分化为造血细胞?需要那些必要的条件?移植后能否进行造血重建?针对上述问题,本研究首先分离人骨髓MSC,建立大规模培养及扩增体系,并对其生物学特性进行检测。然后利用小鼠胎肝基质细胞条件培养液、造血因子,对其进行体外诱导分化,其次以绿色荧光蛋白转染hMSC , 获得稳定表达绿色荧光蛋白的hMSC (hMSC/EGFP)。最后以hMSC 和hMSC/EGFP 为供体细胞,亚致死剂量射线照射后的裸鼠为受体,绿色荧光蛋白作为示踪,通过免疫组化、流式细胞术等方法观察hMSC 的体内造血分化潜能和造血重建能力,结果表明所分离、培养及扩增的hMSC 均一性好,具有多潜能成体干细胞特性。在体外,胎肝基质细胞条件培养液可诱导hMSC 分化为具有造血干/祖细胞特性的细胞。hMSC 能在亚致死剂量射线照射的裸鼠骨髓中植入并分化表型为人CD34+的造血干/祖细胞。
[Abstract]:The bone marrow mesenchymal stem cell (MSC) is a kind of stem cell with multi-directional differentiation potential, which is cultured in vitro and can maintain its genetic character stably for a long time. Hematopoietic stem cell transplantation (HSCT) is an important biological or cellular therapy. Although a good clinical curative effect has been achieved, it is not enough for the source, and cannot be amplified in a large amount in vitro, so that the application of HSCT is still limited in many ways. So, can human bone marrow MSC (hMSCs) differentiate into hematopoietic cells? What are the necessary conditions? Can the hemopoietic reconstruction be performed after transplantation? In view of the above problems, this study first separates human bone marrow MSC, establishes a large-scale culture and amplification system, and detects its biological characteristics. and then the hMSCs were transfected with green fluorescent protein to obtain hMSCs (hMSCs/ EGFP) stably expressing the green fluorescent protein. The hematopoietic differentiation potential and the hematopoietic reconstitution ability of hMSCs were observed by using hMSC and hMSC/ EGFP as the receptor and the green fluorescent protein as the tracer. The results showed that the cells were separated. The hMSCs cultured and amplified have good properties and have the characteristics of multipotent stem cell. In vitro, the fetal liver stromal cell conditioned medium may induce hMSCs to differentiate into cells with hematopoietic stem/ progenitor cell characteristics. hMSCs can be implanted and differentiated into hematopoietic stem/ progenitor cells of human CD34 + in the bone marrow of bare mice irradiated with sublethal dose radiation.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R329.2

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