本文選題：大鼠 + 骨髓間充質干細胞��； 參考：《昆明醫(yī)學院》2011年碩士論文

【摘要】：[目的]：大鼠骨髓間充質干細胞(rabbit mesenchymal stem cells, MSC)分離純化培養(yǎng)后,誘導為成骨細胞、心肌細胞和胰島樣細胞,觀察不同分化階段的細胞形態(tài)、檢測特殊因子或標志物的表達變化。 [方法]：體外分離培養(yǎng)鼠骨髓間充質干細胞,采用文獻中的誘導方法誘導,分別于第3、7、14、21天倒置熒光顯微鏡動態(tài)觀察細胞形態(tài)的變化；并通過RT-PCR技術和酶聯(lián)免疫吸附技術檢測干細胞在誘導不同階段移植特殊因子或標志物的表達變化。 [結果]：MSC能夠正常貼壁成長,抗原表達為CD34-/CD45-/CD73+/CD105+。倒置螢光顯微鏡觀察下細胞形態(tài)發(fā)生明顯變化,RT-PCR檢測結果顯示GATA-4、Nkx2.5、Sarcomericactin和Connexin-43mRNA向誘導成心肌細胞的過程中表達持續(xù)增加,Isulin 1、Isulin 2、CK-19、nestin、Glut2和pdxl mRNA表達在向胰島樣細胞誘導的過程中持續(xù)增加,說明在誘導劑誘導下能夠向誘導為成骨細胞、心肌細胞和胰島樣細胞分化。 [結論]：①建立了GFP標記的RMSC的體外分離、培養(yǎng)及鑒定的技術平臺。②建立了MSC向心肌細胞、成骨細胞和胰島樣細胞分化的誘導體系。⑧RT-PCR結果顯示MSC成功向心肌細胞和胰島樣細胞分化,并分泌相應細胞因子。④酶聯(lián)免疫吸附實驗結果顯示MSC向胰島樣細胞和成骨細胞分化,并且分泌胰島素和堿性磷酸酶。
[Abstract]:[objective]: rat bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured in rabbit mesenchymal stem cells, MSC), then induced into osteoblasts, cardiomyocytes and islet like cells. [methods] Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro, induced by the induction method in literature, and the morphological changes of the cells were observed by inverted fluorescence microscope on day 3, 7, 1421, respectively. RT-PCR and enzyme linked immunosorbent assay (Elisa) were used to detect the expression of specific factors or markers in different stages of stem cell transplantation. [results] CD34-/CD45-/CD73 / CD105 was expressed as CD34-/CD45-/CD73 / CD105 antigen. The results of reverse fluorescent microscopy showed that the expression of Glut2 and pdxl mRNA in islet like cells increased continuously during the process of inducing cardiac myocytes by GATA-4Nkx2.5Nkx2.5A, and the expression of Glut2 and pdxl mRNA increased continuously during the induction of cardiac myocytes, and the expression of Glut2 and pdxl mRNA increased during the induction of Glut2 and pdxl mRNA to islet like cells, and the expression of Glut2 and Glut2 in the islet like cells was increased during the induction of Glut2 and pdxl mRNA to islet like cells. These results indicated that induced osteoblasts, cardiomyocytes and islet like cells could be induced. [conclusion] the in vitro isolation, culture and identification of RMSC labeled by GFP were established in vitro. 2. 2. MSC to cardiomyocytes was established by 1: 1. The differentiation of osteoblasts and islet like cells was induced by .8RT-PCR. The results showed that MSC successfully differentiated into cardiomyocytes and islet like cells. The results of enzyme linked immunosorbent assay (Elisa) showed that MSC differentiated into islet like cells and osteoblasts and secreted insulin and alkaline phosphatase.
【學位授予單位】：昆明醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R329

【參考文獻】

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本文編號：1908122