骨髓间充质干细胞的定向与共培养体系诱导

发布时间：2018-04-28 01:45

本文选题：骨髓间充质干细胞 + 转化　；参考：《青岛大学》2007年硕士论文

【摘要】： 目的通过骨髓间充质干细胞的定向诱导，探讨其成脂、成神经等多分化潜能。利用共培养体系诱导骨髓间充质干细胞向胰岛祖细胞的初步转化，探讨骨髓间充质干细胞在糖尿病治疗领域的价值。方法骨髓间充质干细胞来源于5～10岁儿童胸骨骨髓血(排除造血系统疾病，经得家属同意)。Percoll淋巴细胞分层液分离骨髓间充质干细胞；第3代细胞经CD44抗体、CD45抗体、CD34抗体鉴定；第4代细胞采用含10~(-8)mmol／L地塞米松、10μg／L胰岛素的高糖DMEM(H-DMEM)诱导其向脂肪细胞分化，油红O染色鉴定脂肪细胞。碱性成纤维细胞生长因子(bFGF)诱导其向神经祖细胞分化，经巢蛋白鉴定，继以1％二甲基亚矾(DMSO)的无血清H-DMEM诱导其形成神经网格样结构。胶原酶消化法获取流产胎儿胰腺间充质细胞，，用添加bFGF的无血清低糖DMEM(L-DMEM)使其增殖，然后将胰腺间充质细胞接种于底层已接种骨髓间充质干细胞的Transwell内板(insert)，共培养法诱导骨髓间充质干细胞向胰岛祖细胞的初步转化。建立骨髓间充质干细胞和神经干细胞共培养体系诱导神经干细胞分化为神经元。结果骨髓间充质干细胞体外经诱导可分化为脂肪细胞及神经细胞。胎儿胰腺间充质细胞中具有巢蛋白、CK-19阳性的胰腺干细胞，体外可分化形成胰岛样细胞团。共培养法可诱导骨髓间充质干细胞分化为PDX-1免疫反应阳性细胞。共培养体系能促进神经干细胞的分化。结论Percoll淋巴细胞分离液分离可获取骨髓间充质干细胞，体外诱导可分化为脂肪细胞、神经细胞。胎儿胰腺微环境能诱导骨髓间充质干细胞向胰岛祖细胞的转化。骨髓间充质干细胞分泌的一些物质对神经干细胞的分化具有促进作用。
[Abstract]:Objective to investigate the multiple differentiation potential of bone marrow mesenchymal stem cells (BMSCs). The primary transformation of bone marrow mesenchymal stem cells to islet progenitor cells was induced by co-culture system, and the value of bone marrow mesenchymal stem cells in the treatment of diabetes was discussed. Methods Bone marrow mesenchymal stem cells (BMSCs) derived from sternal bone marrow blood (excluding hematopoietic system diseases) were isolated from bone marrow mesenchymal stem cells (BMSCs) from the sternal bone marrow blood of children aged 5 to 10 years old (excluding hematopoietic system diseases), and the third passage cells were identified by CD44 antibody CD45 antibody and CD34 antibody. The fourth passage cells were induced to differentiate into adipocytes by high glucose DMEMN H-DMEM containing 10 渭 g / L insulin of 10~(-8)mmol/L dexamethasone. The adipocytes were identified by oil red O staining. Basic fibroblast growth factor (bFGF) induced its differentiation into neural progenitor cells, and then the serum-free H-DMEM of 1% DMSOs was used to induce the formation of neural mesh-like structure. Fetal pancreatic mesenchymal cells were obtained by collagenase digestion and proliferated by serum free low-glucose DMEML-DMEM supplemented with bFGF. Then the pancreatic mesenchymal cells were inoculated into the Transwell inner plate of bone marrow mesenchymal stem cells (BMSCs) in the bottom layer, and the primary transformation of BMSCs to islet progenitor cells was induced by co-culture. The co-culture system of bone marrow mesenchymal stem cells and neural stem cells was established to induce neural stem cells to differentiate into neurons. Results Bone marrow mesenchymal stem cells were induced to differentiate into adipocytes and nerve cells in vitro. Pancreatic stem cells positive for nestin and CK-19 were found in fetal pancreatic mesenchymal cells, which could differentiate into islet like cell clusters in vitro. Co-culture can induce bone marrow mesenchymal stem cells to differentiate into PDX-1 immunoreactive cells. Co-culture system can promote the differentiation of neural stem cells. Conclusion Bone marrow mesenchymal stem cells can be obtained by isolating Percoll lymphocytes and can be induced to differentiate into adipocytes and nerve cells in vitro. Fetal pancreatic microenvironment can induce the transformation of bone marrow mesenchymal stem cells to islet progenitor cells. Some substances secreted by bone marrow mesenchymal stem cells promote the differentiation of neural stem cells.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R329

【共引文献】