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醫(yī)學(xué)論文范文:慢性高原病患者骨髓間充質(zhì)干細(xì)胞的體外擴(kuò)增及鑒定
【摘要】 目的 探討慢性高原病(chronic mountain sickness，CMS)患者骨髓間充質(zhì)干細(xì)胞(mesenchymal stem cell，MSCs)的體外培養(yǎng)方法、生物功能及分子生物學(xué)特征。方法 采用密度梯度離心和貼壁篩選法對(duì)15例CMS患者(CMS組)和15例正常人(對(duì)照組)骨髓MSCs進(jìn)行分離培養(yǎng)，于倒置顯微鏡下觀察MSCs形態(tài)，用流式細(xì)胞儀行細(xì)胞表面抗原檢測，檢測MSCs生長情況，測定生長曲線。結(jié)果 采用密度梯度離心法分離骨髓單個(gè)核細(xì)胞(BMMNC)后培養(yǎng)，骨髓MSC均分離培養(yǎng)成功，貼壁細(xì)胞呈梭形;兩組骨髓MSCs均表達(dá)CD29、CD105和CD13，不表達(dá)CD45、CD4、CD8、CD14、CD3、CD34、HLADR和CD20。CMS組MSCs增殖能力高于對(duì)照組(P<0.01)，但傳代率低于對(duì)照組(P<0.05)，對(duì)P3代細(xì)胞生長曲線進(jìn)行觀察發(fā)現(xiàn)CMS骨髓MSCs生長較對(duì)照組活躍。結(jié)論 CMS患者骨髓MSCs在體外可有效分離培養(yǎng)，所培養(yǎng)擴(kuò)增的細(xì)胞成分單一，CMS患者骨髓MSCs具有其特有的生物學(xué)特性。
【關(guān)鍵詞】 慢性高原病 細(xì)胞培養(yǎng) 間充質(zhì)干細(xì)胞 生物學(xué)特性
INVITROAMPLIFICATION AND IDENTIFICATION OFBONE MARROW MESENCHYMAL STEM CELLS DERIVED醫(yī).學(xué)全.在.線網(wǎng)站m.gydjdsj.org.cn
FROM PATIENTS WITH CHRONIC MOUNTAIN SICKNESS
Ji Linhua, Sun Jing, Wang Hongxin, Cui Sen, Li Zhanquan, Ge Rili
( Department of Hematology, Qinghai University Affiliated Hospital)
Abstract Objective To explore the in vitro culture approaches, biological functions and biological molecular characteristics of bone marrow mesenchymal stem cells(MSCs) derived from patients with chronic mountain sickness(CMS). Methods The CMS was diagnosed by the Qinghai CMS Score established by the VI World Congress in 2004.MSCs were separated from bone marrow of 15 CMS patients(CMS group) and 15 healthy adults(control group) after their agreement by density gradient centrifugation and adherent culture in vitro. MSCs were identified according to morphology and cell surface antigen profile. The morphological changes and the proliferation growth curves of MSCs were observed in primary and passage cultures. The surface makers were detected by flow Cytometry(FCM).Results MSCs were successfully isolated from 15 CMS patients and controls. Adherent cells were spindle shape in morphology. It was found that MSCs expressed CD29,CD105 and CD13 in flow Cytometry, but did not express CD45,CD4,CD8,CD14,CD3,CD34,HLADR and CD20.CMSMSCs shared a similar morphologic characteristics and immune phenotypes with MSCs derived from normal bone marrow. The proliferation rate of MSCs in the CMS group was higher than that in the control group (P<0.01), but the passage rate was lower than that in the control group (P<0.05).It was found that the growth curve of the passage 3 cells that the growth of MSCs derived from CMS patients was more active than that of MSCs derived from the controls. Conclusion The bone marrow MSCs of CMS patients can be cultured purely and can extensively proliferate. And they possess specialbiological characteristics.
Key words Chronic mountain sickness Cell culture Mesenchymal stem cell Biological characteristics
