小鼠骨髓多能干细胞建系及其造血重建作用的研究
发布时间：2019-07-08 10:18
【摘要】：在人和动物的个体发育过程中,胚胎和成体组织均存在具有高度自我更新能力和分化潜能的干细胞。干细胞的分离和体外培养为探讨胚胎发生、组织细胞分化、基因表达调控等发育生物学问题提供了理想的模型,也为多种疾病的细胞替代治疗和基因治疗开拓了新的途径。骨髓中存在多种来源及分化方向不同的成体干细胞,例如造血干细胞、间充质干细胞、内皮前体细胞、成血成血管干细胞等,骨髓来源的干细胞是迄今为止研究最为深入并得到广泛应用的成体干细胞。大量的动物移植模型中发现骨髓来源的干细胞具有可塑性,可转化为肝细胞、心肌细胞、骨骼肌细胞、神经细胞、皮肤角质细胞、胰岛细胞,以及肾脏、肺脏、消化道的上皮细胞等,其可塑性涉及横向分化、去分化、细胞融合等机制,但也可能是由于骨髓中存在多能干细胞,能在各种环境信号作用下向不同组织细胞方向分化的结果。如能分离出骨髓中可能存在的多能干细胞,并进行体外培养、建系,将为研究干细胞的增殖分化调控提供良好的技术平台,为细胞治疗、组织工程技术提供极具前景的种子细胞。 造血干细胞移植(HSCT)目前仍面临几个亟待解决的问题,如供体来源不足、造血恢复时间长及移植物抗宿主病(GVHD)等。骨髓来源多能干细胞的一些生物学特性有可能使其成为解决上述问题的良好靶细胞。首先,已有研究分别从人、大鼠和小鼠骨髓中分离出一种在单个细胞水平即能向三个胚层的多种组织细胞分化的多能干细胞,将该种细胞经尾静脉注入NOD/SCID小鼠,可在骨髓、外周血及脾脏检测到由其分化形成的造血细胞,提示骨髓多能干细胞有可能成为HSC的新来源。其次,骨髓多能干细胞不表达MHC-Ⅱ类抗原,低表达或不表达MHC-Ⅰ类抗原,免疫原性低,因此异基因移植不易被受体排斥,引起GVHD的几率减小。第三,骨髓多能干细胞是在MSC培养过程中发现的一类特殊的干细胞,同时具有与MSC类似的部分生物学特征。已有大量研究表明MSC与造血干细胞(HSC)共移植可促进HSC植入和造血恢复,并有抑制移植排斥和减轻GVHD的作用。目前骨髓多能干细胞的研究尚处于起步阶段,有关移植后造血重建及免疫调节方面的作用国内外鲜有报道,尚需进一步探讨。 基于上述考虑,本课题分离培养小鼠骨髓多能干细胞(mMMSC),建立mMMSC细胞系,验证mMMSC跨系跨胚层分化的多向分化潜能,鉴定其各项生物学特性,为体内外实验提供一个良好的靶细胞。进一步建立mMMSC移植的动物模型,明确
文内图片：
图片说明：A:原代培养形成的集落B:D:mMMSC(密度5x102/e耐)E:体外培养的mMMSCmMMSC普通光镜照片
[Abstract]:In the individual development of human and animal, both the embryonic and adult tissues have stem cells with high self-renewal capacity and differentiation potential. The isolation and in vitro culture of stem cells provide an ideal model for the development of development biology such as embryogenesis, cell differentiation, gene expression regulation and the like, and also opens up a new way for cell replacement therapy and gene therapy of various diseases. In the bone marrow, many adult stem cells, such as hematopoietic stem cells, mesenchymal stem cells, endothelial progenitor cells, and blood-derived stem cells, are present in the bone marrow. The stem cells of the bone marrow are the most in-depth and widely used adult stem cells. In a large number of animal transplantation models, the bone marrow-derived stem cells are found to have plasticity, and can be converted into liver cells, cardiac muscle cells, skeletal muscle cells, nerve cells, skin cutin cells, islet cells, and epithelial cells of the kidney, the lung, the digestive tract, and the like, The plasticity relates to mechanisms such as lateral differentiation, dedifferentiation, cell fusion and the like, but it may also be a result of differentiation of different tissue cells under various environmental signals due to the presence of pluripotent stem cells in the bone marrow. If the pluripotent stem cells which may be present in the bone marrow can be separated, and the in vitro culture and construction system are carried out, the invention provides a good technical platform for the regulation and regulation of the proliferation and differentiation of the research stem cells, and provides a promising seed cell for the cell therapy and tissue engineering technology. Hematopoietic stem cell transplantation (HSCT) is still facing a number of problems to be solved, such as insufficient donor origin, long time of hematopoietic recovery and graft-versus-host Disease (GVHD) and the like. Some of the biological characteristics of the bone marrow-derived pluripotent stem cells can make it possible to solve the above-mentioned problems. Good target cells for the problem. First of all, a number of pluripotent stem cells differentiated from the human, rat and mouse bone marrow are isolated from the bone marrow of the human, rat and mouse, and the cells are injected into the NOD/ SCI through the tail vein. In the D mice, the hematopoietic cells formed by the differentiation can be detected in the bone marrow, the peripheral blood and the spleen, suggesting that the bone marrow pluripotent stem cells are likely to be It is a new source of HSC. Second, the bone marrow pluripotent stem cells do not express MHC-II antigens, are low in expression or do not express MHC-I antigens, and are low in immunogenicity, so the allogenic transplantation is not easy to be repelled by the receptor, causing G The probability of VHDs is reduced. Third, the bone marrow pluripotent stem cells are a class of special stem cells that are found in the MSC culture, and are similar to the MSC A large number of studies have shown that the transplantation of MSC and hematopoietic stem cells (HSC) can promote the implantation of HSC and the recovery of hemopoietic stem cells, and inhibit the rejection and reduction of the transplantation. At present, the research of bone marrow pluripotent stem cells is still in the early stage, and there are few reports on the effects of hematopoietic reconstruction and immunoregulation after transplantation. Based on the above considerations, the mouse bone marrow pluripotent stem cell (mMMSC) was isolated and cultured, and the mMMSC cell line was established to verify the multi-directional differentiation potential of the mMMSC cross-system cross-system to identify its biological characteristics. And provides a good target cell for in-vivo experiments.
【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
本文编号：2511517
文内图片：
图片说明：A:原代培养形成的集落B:D:mMMSC(密度5x102/e耐)E:体外培养的mMMSCmMMSC普通光镜照片
[Abstract]:In the individual development of human and animal, both the embryonic and adult tissues have stem cells with high self-renewal capacity and differentiation potential. The isolation and in vitro culture of stem cells provide an ideal model for the development of development biology such as embryogenesis, cell differentiation, gene expression regulation and the like, and also opens up a new way for cell replacement therapy and gene therapy of various diseases. In the bone marrow, many adult stem cells, such as hematopoietic stem cells, mesenchymal stem cells, endothelial progenitor cells, and blood-derived stem cells, are present in the bone marrow. The stem cells of the bone marrow are the most in-depth and widely used adult stem cells. In a large number of animal transplantation models, the bone marrow-derived stem cells are found to have plasticity, and can be converted into liver cells, cardiac muscle cells, skeletal muscle cells, nerve cells, skin cutin cells, islet cells, and epithelial cells of the kidney, the lung, the digestive tract, and the like, The plasticity relates to mechanisms such as lateral differentiation, dedifferentiation, cell fusion and the like, but it may also be a result of differentiation of different tissue cells under various environmental signals due to the presence of pluripotent stem cells in the bone marrow. If the pluripotent stem cells which may be present in the bone marrow can be separated, and the in vitro culture and construction system are carried out, the invention provides a good technical platform for the regulation and regulation of the proliferation and differentiation of the research stem cells, and provides a promising seed cell for the cell therapy and tissue engineering technology. Hematopoietic stem cell transplantation (HSCT) is still facing a number of problems to be solved, such as insufficient donor origin, long time of hematopoietic recovery and graft-versus-host Disease (GVHD) and the like. Some of the biological characteristics of the bone marrow-derived pluripotent stem cells can make it possible to solve the above-mentioned problems. Good target cells for the problem. First of all, a number of pluripotent stem cells differentiated from the human, rat and mouse bone marrow are isolated from the bone marrow of the human, rat and mouse, and the cells are injected into the NOD/ SCI through the tail vein. In the D mice, the hematopoietic cells formed by the differentiation can be detected in the bone marrow, the peripheral blood and the spleen, suggesting that the bone marrow pluripotent stem cells are likely to be It is a new source of HSC. Second, the bone marrow pluripotent stem cells do not express MHC-II antigens, are low in expression or do not express MHC-I antigens, and are low in immunogenicity, so the allogenic transplantation is not easy to be repelled by the receptor, causing G The probability of VHDs is reduced. Third, the bone marrow pluripotent stem cells are a class of special stem cells that are found in the MSC culture, and are similar to the MSC A large number of studies have shown that the transplantation of MSC and hematopoietic stem cells (HSC) can promote the implantation of HSC and the recovery of hemopoietic stem cells, and inhibit the rejection and reduction of the transplantation. At present, the research of bone marrow pluripotent stem cells is still in the early stage, and there are few reports on the effects of hematopoietic reconstruction and immunoregulation after transplantation. Based on the above considerations, the mouse bone marrow pluripotent stem cell (mMMSC) was isolated and cultured, and the mMMSC cell line was established to verify the multi-directional differentiation potential of the mMMSC cross-system cross-system to identify its biological characteristics. And provides a good target cell for in-vivo experiments.
【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
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相关期刊论文 前4条
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