人骨髓间充质干细胞分化潜能的探讨

发布时间：2018-06-01 11:20

本文选题：间充质干细胞 + 人骨髓　；参考：《第二军医大学》2005年博士论文

【摘要】：人骨髓间充质干细胞(human mesenchymal stem cells,hMSCs)是骨髓中的一类与造血干细胞(hematopoietic stem cells,HSCs)不同的,具有向骨、软骨、脂肪、肌肉等多种组织细胞分化能力的成体干细胞,它在体外可以大量增殖,并具有克隆形成的能力。它就是人们很早就知道的骨髓基质细胞(bone marrow stromal cells,BMSCs)。由于其具有多向分化的潜能,在体外又较容易培养扩增,加之具有潜在的细胞治疗价值,故引起了生物学界和医学界的关注。然而,相对于HSCs来说,人们对MSCs生物学特性的认识要滞后得多,就连其表型特征和分化潜能的认识也尚不一致。造成这种情况的主要原因可能有:1)个体发育的不同阶段,MSCs的表型和功能有差异;2)骨髓中存在不同分化等级的MSCs;3)骨髓中不同空间位置的MSCs存在差异;4)不同实验室分离MSCs的方法不同,分离后体外培养的条件也不一样,以致所获得的MSCs也不同;5)用于研究MSCs分化潜能的体外诱导和体内移植的体系不一致。因此,MSCs迄今尚无公认的、高度特异性的分子标记,也没有较为标准的分离和培养方法。Friedenste AJ(1970)和Gronthos S(2003)等人的研究发现体外培养的单个骨髓基质细胞贴壁生长后可以形成克隆,称为集落形成成纤维样细胞(clonogenic precursors,或colony-forming fibroblastic units,CFU-F),一般所说的骨髓间充质干细胞很可能就是其中的某种或某些类型的细胞群体。基于这样一个研究背景,本课题试图直接从人骨髓中分离较为原始的骨髓间充质干细胞,以期对MSCs的基本生物学特性能够有一个比较全面的认识。本文的目的在于:1)建立克隆化的人骨髓间充质干细胞系,并建立适合其生长和增殖的体外培养条件;2)鉴定它的基本生物学特性;3)研究其向外胚层神经细胞和内胚层肝脏细胞分化的潜能。本实验以年龄为17y-65y的正常人骨髓为材料,利用hMSCs贴壁生长和具集落形成能力的特点,得到了多个克隆化的细胞系。对其中的一个可稳定传代的细胞系(hMSCs-2002-3)的基本生物学特性进行了系统的研究,探讨了骨髓hMSCs在体内和体外的分化潜能,以及其在损伤肝和神经系统中的植入和分化命运。所获得的主要结果如下: (1)hMSCs-2002-3具有间充质干细胞基本的生物学特性。选取骨髓细胞原代培养物
[Abstract]:Human bone marrow mesenchymal stem cells (mesenchymal stem cells) are different from hematopoietic stem cells (HSCs) in bone marrow. They have the ability to differentiate into bone, cartilage, fat, muscle and so on. They can proliferate a lot in vitro. And has the ability of clone formation. It is the bone marrow stromal cells known as bone marrow stromal cells and BMSCs. Because it has the potential of multidirectional differentiation, it is easy to be cultured and amplified in vitro, and has the potential value of cell therapy, it has attracted the attention of biology and medicine. However, the understanding of biological characteristics of MSCs is much slower than that of HSCs, and even the understanding of phenotypic characteristics and differentiation potential of MSCs is not consistent. The main reasons for this may be: (1) phenotypic and functional differences in different stages of ontogeny (2) different differentiation grades of MSCs in bone marrow; (4) different laboratory scores of MSCs with different spatial positions in bone marrow Different from MSCs, The culture conditions were different after isolation, so that the MSCs obtained was different from the system used to study the differentiation potential of MSCs in vitro and in vivo. So far, there is no recognized and highly specific molecular marker, and there is no standard method of isolation and culture. Friedenste AJ (1970) and Gronthos Schi2003). It was found that a single bone marrow stromal cell cultured in vitro could form clone after adherent growth. The bone marrow mesenchymal stem cells (BMSCs), known as colony forming fibroblast cells, or colony-forming fibroblastic unitsCF-FU cells, may be one or some of these cell populations. Based on this background, we try to isolate the primitive bone marrow mesenchymal stem cells directly from human bone marrow in order to have a comprehensive understanding of the basic biological characteristics of MSCs. The purpose of this paper is to establish a cloned human bone marrow mesenchymal stem cell line. Its basic biological characteristics were identified. (3) the potential of its differentiation into ectodermal nerve cells and endoderm liver cells was studied. In this study, a number of cloned cell lines were obtained from normal human bone marrow of age 17y-65y, using the characteristics of hMSCs adherent growth and colony forming ability. The basic biological characteristics of one of the stable passaged cell lines hMSCs-2002-3 were systematically studied. The differentiation potential of bone marrow hMSCs in vivo and in vitro and its fate of implantation and differentiation in injured liver and nervous system were discussed. The main results obtained are as follows: HMSCs-2002-3 has the basic biological characteristics of mesenchymal stem cells. Selection of primary culture of bone marrow cells
【学位授予单位】：第二军医大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R329.2

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