【摘要】：在人和動物的個體發(fā)育過程中,胚胎和成體組織均存在具有高度自我更新能力和分化潛能的干細(xì)胞。干細(xì)胞的分離和體外培養(yǎng)為探討胚胎發(fā)生、組織細(xì)胞分化、基因表達(dá)調(diào)控等發(fā)育生物學(xué)問題提供了理想的模型,也為多種疾病的細(xì)胞替代治療和基因治療開拓了新的途徑。骨髓中存在多種來源及分化方向不同的成體干細(xì)胞,例如造血干細(xì)胞、間充質(zhì)干細(xì)胞、內(nèi)皮前體細(xì)胞、成血成血管干細(xì)胞等,骨髓來源的干細(xì)胞是迄今為止研究最為深入并得到廣泛應(yīng)用的成體干細(xì)胞。大量的動物移植模型中發(fā)現(xiàn)骨髓來源的干細(xì)胞具有可塑性,可轉(zhuǎn)化為肝細(xì)胞、心肌細(xì)胞、骨骼肌細(xì)胞、神經(jīng)細(xì)胞、皮膚角質(zhì)細(xì)胞、胰島細(xì)胞,以及腎臟、肺臟、消化道的上皮細(xì)胞等,其可塑性涉及橫向分化、去分化、細(xì)胞融合等機(jī)制,但也可能是由于骨髓中存在多能干細(xì)胞,能在各種環(huán)境信號作用下向不同組織細(xì)胞方向分化的結(jié)果。如能分離出骨髓中可能存在的多能干細(xì)胞,并進(jìn)行體外培養(yǎng)、建系,將為研究干細(xì)胞的增殖分化調(diào)控提供良好的技術(shù)平臺,為細(xì)胞治療、組織工程技術(shù)提供極具前景的種子細(xì)胞。 造血干細(xì)胞移植(HSCT)目前仍面臨幾個亟待解決的問題,如供體來源不足、造血恢復(fù)時間長及移植物抗宿主病(GVHD)等。骨髓來源多能干細(xì)胞的一些生物學(xué)特性有可能使其成為解決上述問題的良好靶細(xì)胞。首先,已有研究分別從人、大鼠和小鼠骨髓中分離出一種在單個細(xì)胞水平即能向三個胚層的多種組織細(xì)胞分化的多能干細(xì)胞,將該種細(xì)胞經(jīng)尾靜脈注入NOD/SCID小鼠,可在骨髓、外周血及脾臟檢測到由其分化形成的造血細(xì)胞,提示骨髓多能干細(xì)胞有可能成為HSC的新來源。其次,骨髓多能干細(xì)胞不表達(dá)MHC-Ⅱ類抗原,低表達(dá)或不表達(dá)MHC-Ⅰ類抗原,免疫原性低,因此異基因移植不易被受體排斥,引起GVHD的幾率減小。第三,骨髓多能干細(xì)胞是在MSC培養(yǎng)過程中發(fā)現(xiàn)的一類特殊的干細(xì)胞,同時具有與MSC類似的部分生物學(xué)特征。已有大量研究表明MSC與造血干細(xì)胞(HSC)共移植可促進(jìn)HSC植入和造血恢復(fù),并有抑制移植排斥和減輕GVHD的作用。目前骨髓多能干細(xì)胞的研究尚處于起步階段,有關(guān)移植后造血重建及免疫調(diào)節(jié)方面的作用國內(nèi)外鮮有報道,尚需進(jìn)一步探討。 基于上述考慮,本課題分離培養(yǎng)小鼠骨髓多能干細(xì)胞(mMMSC),建立mMMSC細(xì)胞系,驗證mMMSC跨系跨胚層分化的多向分化潛能,鑒定其各項生物學(xué)特性,為體內(nèi)外實驗提供一個良好的靶細(xì)胞。進(jìn)一步建立mMMSC移植的動物模型,明確
文內(nèi)圖片：
圖片說明：A:原代培養(yǎng)形成的集落B:D:mMMSC(密度5x102/e耐)E:體外培養(yǎng)的mMMSCmMMSC普通光鏡照片
[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.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2005
【分類號】：R329

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