本文選題：骨髓間充質(zhì)干細胞 + 轉(zhuǎn)化�。� 參考：《青島大學(xué)》2007年碩士論文

【摘要】： 目的通過骨髓間充質(zhì)干細胞的定向誘導(dǎo)，探討其成脂、成神經(jīng)等多分化潛能。利用共培養(yǎng)體系誘導(dǎo)骨髓間充質(zhì)干細胞向胰島祖細胞的初步轉(zhuǎn)化，探討骨髓間充質(zhì)干細胞在糖尿病治療領(lǐng)域的價值。 方法骨髓間充質(zhì)干細胞來源于5～10歲兒童胸骨骨髓血(排除造血系統(tǒng)疾病，經(jīng)得家屬同意)。Percoll淋巴細胞分層液分離骨髓間充質(zhì)干細胞；第3代細胞經(jīng)CD44抗體、CD45抗體、CD34抗體鑒定；第4代細胞采用含10~(-8)mmol／L地塞米松、10μg／L胰島素的高糖DMEM(H-DMEM)誘導(dǎo)其向脂肪細胞分化，油紅O染色鑒定脂肪細胞。堿性成纖維細胞生長因子(bFGF)誘導(dǎo)其向神經(jīng)祖細胞分化，經(jīng)巢蛋白鑒定，繼以1％二甲基亞礬(DMSO)的無血清H-DMEM誘導(dǎo)其形成神經(jīng)網(wǎng)格樣結(jié)構(gòu)。 膠原酶消化法獲取流產(chǎn)胎兒胰腺間充質(zhì)細胞，，用添加bFGF的無血清低糖DMEM(L-DMEM)使其增殖，然后將胰腺間充質(zhì)細胞接種于底層已接種骨髓間充質(zhì)干細胞的Transwell內(nèi)板(insert)，共培養(yǎng)法誘導(dǎo)骨髓間充質(zhì)干細胞向胰島祖細胞的初步轉(zhuǎn)化。 建立骨髓間充質(zhì)干細胞和神經(jīng)干細胞共培養(yǎng)體系誘導(dǎo)神經(jīng)干細胞分化為神經(jīng)元。 結(jié)果骨髓間充質(zhì)干細胞體外經(jīng)誘導(dǎo)可分化為脂肪細胞及神經(jīng)細胞。胎兒胰腺間充質(zhì)細胞中具有巢蛋白、CK-19陽性的胰腺干細胞，體外可分化形成胰島樣細胞團。共培養(yǎng)法可誘導(dǎo)骨髓間充質(zhì)干細胞分化為PDX-1免疫反應(yīng)陽性細胞。共培養(yǎng)體系能促進神經(jīng)干細胞的分化。 結(jié)論Percoll淋巴細胞分離液分離可獲取骨髓間充質(zhì)干細胞，體外誘導(dǎo)可分化為脂肪細胞、神經(jīng)細胞。胎兒胰腺微環(huán)境能誘導(dǎo)骨髓間充質(zhì)干細胞向胰島祖細胞的轉(zhuǎn)化。骨髓間充質(zhì)干細胞分泌的一些物質(zhì)對神經(jīng)干細胞的分化具有促進作用。
[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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R329

【共引文獻】

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