發(fā)布時(shí)間：2018-05-29 06:09

  本文選題：骨髓 + 間質(zhì)干細(xì)胞�。� 參考：《中國(guó)組織工程研究》2017年09期

【摘要】：背景:如何能簡(jiǎn)便高效地獲得均質(zhì)性的骨髓間充質(zhì)干細(xì)胞群是軟骨組織工程研究的重點(diǎn)。目的:分離培養(yǎng)大鼠骨髓間充質(zhì)干細(xì)胞,觀察其生物學(xué)特性。方法:抽取30只SD大鼠股骨以及脛骨骨髓,采用全骨髓貼壁培養(yǎng)法體外分離培養(yǎng)骨髓間充質(zhì)干細(xì)胞,流式細(xì)胞儀測(cè)定細(xì)胞表面標(biāo)記物的表達(dá),細(xì)胞計(jì)數(shù)法觀察細(xì)胞生長(zhǎng)增殖能力,MTT法檢測(cè)細(xì)胞存活率。結(jié)果與結(jié)論:(1)剛分離培養(yǎng)的細(xì)胞呈圓形、橢圓形,部分細(xì)胞呈三角形,接種12-15 d后,細(xì)胞開(kāi)始融合,傳代后細(xì)胞增殖速度加快,傳代2 h后細(xì)胞均勻分布在瓶底;(2)隨著傳代次數(shù)的增加,細(xì)胞增殖能力出現(xiàn)下降趨勢(shì),第1-5代細(xì)胞存活率均超過(guò)95%,顯著高于第6代和第7代(P0.05);(3)骨髓間充質(zhì)干細(xì)胞表面CD34呈陰性表達(dá),CD44呈陽(yáng)性表達(dá);(4)結(jié)果表明,采用全骨髓貼壁法可有效分離大鼠骨髓間充質(zhì)干細(xì)胞,獲得的細(xì)胞生長(zhǎng)穩(wěn)定,增殖能力強(qiáng),可選用前5代細(xì)胞作為軟骨組織工程的種子細(xì)胞。
[Abstract]:Background: how to obtain homogeneous bone marrow mesenchymal stem cells (BMSCs) is the focus of cartilage tissue engineering. Objective: to isolate and culture rat bone marrow mesenchymal stem cells (BMSCs) and observe their biological characteristics. Methods: bone marrow of femur and tibia of 30 Sprague-Dawley rats were extracted, bone marrow mesenchymal stem cells were isolated and cultured by whole bone marrow adherent culture in vitro, and the expression of cell surface markers was measured by flow cytometry. Cell proliferation was observed by cell count method and cell survival rate was determined by MTT assay. Results and conclusion: the newly isolated and cultured cells were round, elliptical and triangular. After 12-15 days of inoculation, the cells began to fuse, and the proliferation rate of the cells increased after passage. After 2 h of passage, the cells distributed evenly at the bottom of the bottle.) with the increase of passage times, the proliferative ability of the cells decreased. The survival rate of the 1-5 passage cells exceeded 95%, which was significantly higher than that of the 6th and 7th generation (P0.05 + 3). The results showed that the bone marrow mesenchymal stem cells (BMSCs) could be effectively isolated by whole-bone marrow adherent method, and the positive expression of CD44 on bone marrow mesenchymal stem cells (BMSCs) was negative. The obtained cells can be used as seed cells for cartilage tissue engineering because of their stable growth and strong proliferative ability.
【作者單位】： 江西醫(yī)學(xué)高等專科學(xué)校;南昌大學(xué)醫(yī)學(xué)部;
【分類號(hào)】：R318.08;R68

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