本文選題：基因 + 骨髓間充質(zhì)干細(xì)胞　； 參考：《蘭州大學(xué)》2009年碩士論文

【摘要】： 目的 本研究分離培養(yǎng)骨髓源間充質(zhì)干細(xì)胞(mesenchymal stem cells,MSCs),檢測survivin基因在其中的表達(dá)情況,并分別通過紫外線照射及粒細(xì)胞集落刺激因子刺激后,觀察MSCs中survivin基因的表達(dá)變化,探討其意義。 方法 應(yīng)用Ficoll密度梯度離心法分離骨髓間充質(zhì)干細(xì)胞,LG-DMEM培養(yǎng)基,15%FBS培養(yǎng),培養(yǎng)過程中繪制細(xì)胞生長曲線,流式細(xì)胞術(shù)檢測其特異性表面標(biāo)志,并用地塞米松、胰島素、吲哚美辛誘導(dǎo)成脂,21天后油紅O染色照相,采用半定量RT-PCR法檢測MSCs中survivin基因在轉(zhuǎn)錄水平上的表達(dá)情況。并通過紫外線照射、G-CSF刺激等方法處理后,采用半定量RT-PCR法檢測其中survivin基因表達(dá)的變化。 結(jié)果 原代和傳代培養(yǎng)MSCs均保持較強(qiáng)的增殖能力,細(xì)胞生長曲線提示第7天傳代細(xì)胞進(jìn)入平臺(tái)期,流式細(xì)胞儀檢測示CD83、CD45、組織相容性抗原HLA-DR及干細(xì)胞表面標(biāo)志CD34無表達(dá)或極低表達(dá),而CD44、CD13、CD71高表達(dá),經(jīng)地塞米松等誘導(dǎo)后MSCs向脂肪細(xì)胞分化,RT-PCR結(jié)果顯示正常人骨髓間充質(zhì)干細(xì)胞中有survivin基因表達(dá),并且分別經(jīng)過紫外線照射、G-CSF處理后人骨髓間充質(zhì)干細(xì)胞中survivin基因的表達(dá)量有不同程度的提高。 結(jié)論 Survivin基因不是癌細(xì)胞特異性抗凋亡蛋白,在正常人骨髓間充質(zhì)干細(xì)胞中有表達(dá),并且通過紫外線照射、G-CSF刺激等處理后其表達(dá)量有不同程度的提高,提示它可能參與了間充質(zhì)干細(xì)胞的生物學(xué)行為并影響著間充質(zhì)干細(xì)胞存活。
[Abstract]:PurposeIn this study, mesenchymal stem cells were isolated from bone marrow mesenchymal stem cells and the expression of survivin gene was detected. The changes of survivin gene expression in MSCs were observed after ultraviolet irradiation and granulocyte colony stimulating factor stimulation.MethodBone marrow mesenchymal stem cells were isolated from LG-DMEM medium by Ficoll density gradient centrifugation and cultured in 15s. The cell growth curve was drawn and the specific surface markers were detected by flow cytometry.Indomethacin was induced into lipids for 21 days and oil red O staining was used to detect the expression of survivin gene at the transcriptional level in MSCs by semi-quantitative RT-PCR assay.The changes of survivin gene expression were detected by semi-quantitative RT-PCR after treated by ultraviolet irradiation and G-CSF stimulation.ResultThe cell growth curve indicated that the passage cells entered the plateau phase on the 7th day. Flow cytometry showed that CD83 + CD45, histocompatibility antigen HLA-DR and stem cell surface marker CD34 had no or very low expression.However, CD44, CD13, CD71 were highly expressed. RT-PCR results of MSCs differentiation into adipocytes showed that survivin gene was expressed in normal human bone marrow mesenchymal stem cells.The expression of survivin gene in human bone marrow mesenchymal stem cells was increased by UV irradiation.ConclusionSurvivin gene is not a specific anti-apoptotic protein of cancer cells. It is expressed in normal human bone marrow mesenchymal stem cells.It may be involved in the biological behavior of mesenchymal stem cells and affect the survival of mesenchymal stem cells.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R329

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 何任紅;范建中;吳紅瑛;李德潔;畢迎立;;次聲對(duì)大鼠骨髓間充質(zhì)干細(xì)胞分泌存活素能力影響的研究[J];中國康復(fù)醫(yī)學(xué)雜志;2013年07期

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本文編號(hào)：1737395