人血液血管细胞生成素对骨髓基质细胞生物学作用的实验研究

发布时间：2018-10-20 17:35

【摘要】：本文研究了小鼠骨髓基质细胞的生物学特性,探索了骨髓造血微环境的特征;还研究了基因工程重组的人血液血管细胞生成素对骨髓基质细胞的生物学活性的影响;并以人血液血管细胞生成素基因修饰骨髓基质细胞,用于体外造血的支持。 第一章为了研究骨髓基质细胞(间充质干细胞)的生物学性状和多系分化潜能,我们取Balb/c 小鼠骨髓单个核细胞在低糖的培养液中培养出贴壁生长的细胞,进行形态学观察、细胞周期和免疫表型分析;在不同的细胞因子作用下诱导向成骨细胞、软骨细胞和脂肪细胞分化,并检测诱导后细胞相应的基因表达。结果发现,小鼠骨髓间充质干细胞贴壁生长后形态较均一,增殖能力随着传代逐渐增强,但从第8 代后增殖能力明显减退。细胞表达CD29, CD38, CD44,CD106 等抗原,但CD34 和H-2k 表达阴性。在不同的诱导培养体系里间充质干细胞能分化为成骨细胞、软骨细胞和脂肪细胞,相应细胞的骨钙蛋白基因、Ⅱ型胶原基因和脂蛋白脂酶基因表达都明显增强。 第二章由于血液血管细胞生成素是一种新近被认识的细胞因子,其诸多生物学活性,包括对骨髓基质细胞的作用,还有待于深入探讨。因此,我们研究了基因工程重组的人血液血管细胞生成素(rhHAPO)对原代骨髓基质细胞的增殖和成纤维集落形成单位的影响;对骨髓基质细胞系黏附和增殖及其细胞表面黏附分子的作用;此外,还利用RT-PCR 研究了骨髓基质细胞的细胞因子基因表达情况。结果表明,rhHAPO 通过上调基质细胞的S 期细胞比例,促进骨髓基质细胞的增殖;通过上调VCAM 和ICAM 等细胞表面黏附分子,促进骨髓基质细胞的总黏附能力;而骨髓基质细胞除了表达M-CSF 和c-mpl 等因子基因外,并不表达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，

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