本文關(guān)鍵詞：骨髓間充質(zhì)干細(xì)胞微泡生物學(xué)特性及其促進(jìn)造血干細(xì)胞體外擴(kuò)增作用的研究　出處：《中國(guó)實(shí)驗(yàn)血液學(xué)雜志》2017年04期 　論文類型：期刊論文

  更多相關(guān)文章： 間充質(zhì)干細(xì)胞 微泡 造血干細(xì)胞

【摘要】：目的:探討骨髓間充質(zhì)干細(xì)胞微泡的生物學(xué)特性及其對(duì)造血干細(xì)胞體外擴(kuò)增的作用。方法:用多步差速離心法分離提純骨髓間充質(zhì)干細(xì)胞(MSC)培養(yǎng)上清中的微泡(MV),采用樣本負(fù)染的方法在電子顯微鏡下觀察微泡的形態(tài)特征;用Micro-BCA法測(cè)定其蛋白含量;用流式細(xì)胞術(shù)分析微泡表面標(biāo)志;液體培養(yǎng)動(dòng)員后外周血造血干細(xì)胞實(shí)驗(yàn)分為兩組,在相同培養(yǎng)體系下,給微泡組加入50μl微泡,對(duì)照組加入等體積PBS;采用細(xì)胞計(jì)數(shù)觀察細(xì)胞數(shù)目的變化,應(yīng)用流式細(xì)胞術(shù)動(dòng)態(tài)監(jiān)測(cè)造血干細(xì)胞表面標(biāo)志的變化,細(xì)胞集落培養(yǎng)法觀測(cè)與微泡共培養(yǎng)后造血干細(xì)胞在體外的功能變化。結(jié)果:間充質(zhì)干細(xì)胞來源的微泡是直徑在20-100 nm之間的類圓形囊泡,提取后的微泡濃度約為200μg/ml;在間充質(zhì)干細(xì)胞微泡中CD63表達(dá)率為96.0%,CD44表達(dá)率為50.2%,而HLA-DR,CD34,CD29,CD73等表達(dá)均為陰性;微泡與GPBM NC共培養(yǎng)2 d后,微泡組細(xì)胞數(shù)是對(duì)照組的1.49±0.15倍(P0.05),CD34~+細(xì)胞數(shù)(3.93±0.60)×10~4是對(duì)照組(2.30±0.64)×10~4的1.76±0.30倍;4 d時(shí)微泡組細(xì)胞數(shù)(10.19±0.65)×10~6是實(shí)驗(yàn)組細(xì)胞數(shù)(4.67±0.70)×10~6的2.20±0.24倍(P0.05),微泡組CD34~+細(xì)胞數(shù)(7.82±0.41)×10~4是對(duì)照組(4.03±0.35)×10~4的1.95±0.20倍。結(jié)論:通過多步差速離心法能夠成功從MSC上清中提取微泡,微泡在體外對(duì)造血干細(xì)胞具有促進(jìn)增殖的作用。
[Abstract]:Objective: To investigate the biological characteristics of bone marrow mesenchymal stem cells and microbubbles on hematopoietic stem cell proliferation in vitro. Methods: the separation and purification of bone marrow mesenchymal stem cells by multi-step differential centrifugation (MSC) in the supernatants of microbubbles (MV) method, using negative staining electron microscopy in the sample under the observation of the micro morphological characteristics of bubble; Determination of protein content by Micro-BCA method; using flow cytometric analysis of microbubble surface markers; liquid culture after mobilization of peripheral blood stem cells were divided into two groups, in the same culture system, to join the microbubble group 50 L microbubbles, the control group added volume PBS cells were observed by cell counting; number of changes, changes of stem cell surface markers by flow cytometry and dynamic monitoring of hematopoietic cell colony culture method, observation and microbubble co cultured hematopoietic stem cells change function in vitro. Results: mesenchymal stem cell source 寰場(chǎng)鏄洿寰勫湪20-100 nm涔嬮棿鐨勭被鍦嗗艦鍥婃場(chǎng),鎻愬彇鍚庣殑寰場(chǎng)嫻撳害綰︿負(fù)200渭g/ml;鍦ㄩ棿鍏呰川騫茬粏鑳?yōu)寰场涓瑿D63琛ㄨ揪鐜囦負(fù)96.0%,CD44琛ㄨ揪鐜囦負(fù)50.2%,鑰孒LA-DR,CD34,CD29,CD73絳夎〃杈懼潎涓洪槾鎬，

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