【摘要】：目的：本實(shí)驗(yàn)旨在通過陽離子脂質(zhì)體包埋BMP-2, TGF-β3雙基因修飾質(zhì)粒轉(zhuǎn)染骨髓間充質(zhì)干細(xì)胞,觀察BMP2及TGFβ3在細(xì)胞內(nèi)表達(dá)及對細(xì)胞增殖活性的影響,為下一步研究雙基因表達(dá)載體對間充質(zhì)干細(xì)胞成骨分化影響奠定基礎(chǔ)。 方法：全貼壁法培養(yǎng)兔骨髓間充質(zhì)干細(xì)胞,進(jìn)行原代和傳代培養(yǎng),取P2代細(xì)胞進(jìn)行轉(zhuǎn)染,分別將pIRES/BMP-2、 pIRES/TGFp3、 pIRES/BMP2-TGFp3及空質(zhì)粒轉(zhuǎn)染入干細(xì)胞,并設(shè)置空白對照組,轉(zhuǎn)染兩天后RT-PCR檢測各組BMP2及TGFβ3的表達(dá)情況,繪制細(xì)胞生長曲線,計(jì)算細(xì)胞倍增時間,比較各組干細(xì)胞的增殖情況。 結(jié)果：pIRES/BMP2-TGFβ3轉(zhuǎn)染組BMP2及TGFβ3mRNA含量高于單基因組及空白對照組(p0.05),雙基因均可獲得穩(wěn)定表達(dá)；雙基因組細(xì)胞倍增時間較單一組及空白對照組明顯縮短(p0.05),雙基因組具有良好的促進(jìn)BMCs增殖作用。 結(jié)論：雙基因修飾質(zhì)粒轉(zhuǎn)染干細(xì)胞可以再細(xì)胞內(nèi)獲得穩(wěn)定的表達(dá),并具有良好的協(xié)同促干細(xì)胞增殖活性,為進(jìn)一步研究雙基因質(zhì)粒對誘導(dǎo)間充質(zhì)干細(xì)胞成骨的影響及兩者之間的協(xié)同效應(yīng)提供理論基礎(chǔ)。
[Abstract]:Aim: to investigate the expression of BMP2 and TGF 尾 3 in bone marrow mesenchymal stem cells (BMSCs) transfected with cationic liposome BMP-2, TGF- 尾 3 modified plasmid, and to observe the effect of BMP2 and TGF 尾 3 on the proliferation of bone marrow mesenchymal stem cells. To lay a foundation for the further study of the effect of double gene expression vector on osteogenic differentiation of mesenchymal stem cells. Methods: rabbit bone marrow mesenchymal stem cells (BMSCs) were cultured by whole adherent method, primary and passage culture, P2 generation cells were transfected, pIRES/BMP-2, pIRES/TGFp3, pIRES/BMP2-TGFp3 and empty plasmids were transfected into stem cells, and blank control group was set up. After two days of transfection, RT-PCR was used to detect the expression of BMP2 and TGF 尾 3 in each group, cell growth curve was drawn, cell doubling time was calculated, and the proliferation of stem cells in each group was compared. Results: the contents of BMP2 and TGF 尾 3mRNA in pIRES/BMP2-TGF 尾 3 transfected group were higher than those in single genome and blank control group (p0. 05). The doubling time of double genome cells was significantly shorter than that of single group and blank control group (p0. 05). The double genome could promote the proliferation of BMCs. Conclusion: transfection of double gene modified plasmids into stem cells can obtain stable expression in the cells and has good synergistic effect on the proliferation of stem cells. It provides a theoretical basis for the further study of the effects of double gene plasmids on the osteogenesis of mesenchymal stem cells and their synergistic effects.
【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】：2012
【分類號】：R329

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