【摘要】： 研究目的：通過對(duì)照研究，用形態(tài)學(xué)，免疫組織化學(xué)方法分析TGF-β1在人骨髓間充質(zhì)干細(xì)胞(bone marrow-derived Mesenchymal stem cells，骨髓MSCs)體外定向分化為成纖維細(xì)胞(fibroblast)過程中的作用，并比較TGF-β1在正常人和慢性骨髓增殖性疾病(chronic myeloproliferative disorders，CMPD)病人的骨髓間充質(zhì)干細(xì)胞分化為成纖維細(xì)胞過程中的表達(dá)差異。方法：密度梯度離心法分離正常人和慢性骨髓增殖性疾病病人骨髓液中的間充質(zhì)干細(xì)胞，用含10％胎牛血清(fetal calf serum，F(xiàn)CS)的DMEM低糖雙抗培養(yǎng)液，，在37℃，5％CO_2培養(yǎng)箱中培養(yǎng)，0.25％胰蛋白酶+0.02％EDTA消化傳代；5ng／ml的TGF-β1誘導(dǎo)分化骨髓MSCs，并作空白對(duì)照試驗(yàn)，分別用流式細(xì)胞分析細(xì)胞表面抗原，改良Masson三色染色法初步定性分析膠原蛋白的表達(dá)，免疫組織化學(xué)分析細(xì)胞基質(zhì)蛋白屬性。結(jié)果：聯(lián)合應(yīng)用密度梯度離心法和貼壁法能獲得純度較高，均一性較強(qiáng)的骨髓MSCs同源細(xì)胞，5ng／mlTGF-β1在骨髓間充質(zhì)干細(xì)胞分化培養(yǎng)過程中，能促進(jìn)MSCs表達(dá)膠原蛋白，波形蛋白。同等條件下，慢性骨髓增殖性疾病的MSCs膠原蛋白和波形蛋白表達(dá)高于正常人，細(xì)胞角蛋白的表達(dá)在各組中均低。討論：通過密度梯度離心法結(jié)合貼壁法可以獲得純度較高的骨髓 間充質(zhì)干細(xì)胞，呈梭形、三角形和多角形。表達(dá)CD29、CD44、CD166，不表達(dá)CD14、CD34、HLA-DR、CD45。在骨髓MSCs分化為成纖維細(xì)胞過程中，TGF-β1有促進(jìn)分化的作用。同等條件下CMPD患者骨髓間充質(zhì)干細(xì)胞較正常人的易于表達(dá)膠原蛋白和波形蛋白。
[Abstract]:Objective: to analyze the role of TGF- 尾 1 in the differentiation of human bone marrow mesenchymal stem cells (bone marrow-derived Mesenchymal stem cells, bone marrow MSCs) into fibroblasts (fibroblast) in vitro. The expression of TGF- 尾 1 in bone marrow mesenchymal stem cells (MSCs) differentiated into fibroblasts was compared between normal subjects and patients with chronic myeloproliferation disease (chronic myeloproliferative disorders,CMPD). Methods: mesenchymal stem cells were isolated from bone marrow fluid of normal subjects and patients with chronic myeloproliferation diseases by density gradient centrifugation. DMEM low glucose double antibody medium containing 10% fetal bovine serum (fetal calf serum,FCS was used at 37 鈩

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