發(fā)布時(shí)間：2018-10-20 17:35

【摘要】：本文研究了小鼠骨髓基質(zhì)細(xì)胞的生物學(xué)特性,探索了骨髓造血微環(huán)境的特征;還研究了基因工程重組的人血液血管細(xì)胞生成素對(duì)骨髓基質(zhì)細(xì)胞的生物學(xué)活性的影響;并以人血液血管細(xì)胞生成素基因修飾骨髓基質(zhì)細(xì)胞,用于體外造血的支持。 第一章為了研究骨髓基質(zhì)細(xì)胞(間充質(zhì)干細(xì)胞)的生物學(xué)性狀和多系分化潛能,我們?nèi)alb/c 小鼠骨髓單個(gè)核細(xì)胞在低糖的培養(yǎng)液中培養(yǎng)出貼壁生長(zhǎng)的細(xì)胞,進(jìn)行形態(tài)學(xué)觀察、細(xì)胞周期和免疫表型分析;在不同的細(xì)胞因子作用下誘導(dǎo)向成骨細(xì)胞、軟骨細(xì)胞和脂肪細(xì)胞分化,并檢測(cè)誘導(dǎo)后細(xì)胞相應(yīng)的基因表達(dá)。結(jié)果發(fā)現(xiàn),小鼠骨髓間充質(zhì)干細(xì)胞貼壁生長(zhǎng)后形態(tài)較均一,增殖能力隨著傳代逐漸增強(qiáng),但從第8 代后增殖能力明顯減退。細(xì)胞表達(dá)CD29, CD38, CD44,CD106 等抗原,但CD34 和H-2k 表達(dá)陰性。在不同的誘導(dǎo)培養(yǎng)體系里間充質(zhì)干細(xì)胞能分化為成骨細(xì)胞、軟骨細(xì)胞和脂肪細(xì)胞,相應(yīng)細(xì)胞的骨鈣蛋白基因、Ⅱ型膠原基因和脂蛋白脂酶基因表達(dá)都明顯增強(qiáng)。 第二章由于血液血管細(xì)胞生成素是一種新近被認(rèn)識(shí)的細(xì)胞因子,其諸多生物學(xué)活性,包括對(duì)骨髓基質(zhì)細(xì)胞的作用,還有待于深入探討。因此,我們研究了基因工程重組的人血液血管細(xì)胞生成素(rhHAPO)對(duì)原代骨髓基質(zhì)細(xì)胞的增殖和成纖維集落形成單位的影響;對(duì)骨髓基質(zhì)細(xì)胞系黏附和增殖及其細(xì)胞表面黏附分子的作用;此外,還利用RT-PCR 研究了骨髓基質(zhì)細(xì)胞的細(xì)胞因子基因表達(dá)情況。結(jié)果表明,rhHAPO 通過上調(diào)基質(zhì)細(xì)胞的S 期細(xì)胞比例,促進(jìn)骨髓基質(zhì)細(xì)胞的增殖;通過上調(diào)VCAM 和ICAM 等細(xì)胞表面黏附分子,促進(jìn)骨髓基質(zhì)細(xì)胞的總黏附能力;而骨髓基質(zhì)細(xì)胞除了表達(dá)M-CSF 和c-mpl 等因子基因外,并不表達(dá)HAPO 和
[Abstract]:The biological characteristics of mouse bone marrow stromal cells and the characteristics of hematopoietic microenvironment in bone marrow were studied, and the effects of recombinant human blood angiopoietin on the biological activity of bone marrow stromal cells were also studied. Bone marrow stromal cells were modified with human blood angiopoietin gene to support hematopoiesis in vitro. In chapter 1, in order to study the biological characteristics and multiline differentiation potential of bone marrow stromal cells (MSCs), bone marrow mononuclear cells from Balb/c mice were cultured in low glucose medium to observe the morphology. Cell cycle and immunophenotypic analysis, differentiation into osteoblasts, chondrocytes and adipocytes induced by different cytokines, and the corresponding gene expression were detected. The results showed that the morphology of mouse bone marrow mesenchymal stem cells was uniform after adherent growth, and the proliferative ability increased gradually with passage, but decreased obviously from the 8th generation. The cells expressed CD29, CD38, CD44,CD106 and other antigens, but negative expression of CD34 and H-2k. The expression of osteocalcin gene, type 鈪，

本文編號(hào)：2283894