本文關(guān)鍵詞：骨髓間充質(zhì)干細胞微泡生物學(xué)特性及其促進造血干細胞體外擴增作用的研究　出處：《中國實驗血液學(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)干細胞 微泡 造血干細胞

【摘要】：目的:探討骨髓間充質(zhì)干細胞微泡的生物學(xué)特性及其對造血干細胞體外擴增的作用。方法:用多步差速離心法分離提純骨髓間充質(zhì)干細胞(MSC)培養(yǎng)上清中的微泡(MV),采用樣本負染的方法在電子顯微鏡下觀察微泡的形態(tài)特征;用Micro-BCA法測定其蛋白含量;用流式細胞術(shù)分析微泡表面標志;液體培養(yǎng)動員后外周血造血干細胞實驗分為兩組,在相同培養(yǎng)體系下,給微泡組加入50μl微泡,對照組加入等體積PBS;采用細胞計數(shù)觀察細胞數(shù)目的變化,應(yīng)用流式細胞術(shù)動態(tài)監(jiān)測造血干細胞表面標志的變化,細胞集落培養(yǎng)法觀測與微泡共培養(yǎng)后造血干細胞在體外的功能變化。結(jié)果:間充質(zhì)干細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)干細胞微泡中CD63表達率為96.0%,CD44表達率為50.2%,而HLA-DR,CD34,CD29,CD73等表達均為陰性;微泡與GPBM NC共培養(yǎng)2 d后,微泡組細胞數(shù)是對照組的1.49±0.15倍(P0.05),CD34~+細胞數(shù)(3.93±0.60)×10~4是對照組(2.30±0.64)×10~4的1.76±0.30倍;4 d時微泡組細胞數(shù)(10.19±0.65)×10~6是實驗組細胞數(shù)(4.67±0.70)×10~6的2.20±0.24倍(P0.05),微泡組CD34~+細胞數(shù)(7.82±0.41)×10~4是對照組(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,微泡在體外對造血干細胞具有促進增殖的作用。
[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場鏄洿寰勫湪20-100 nm涔嬮棿鐨勭被鍦嗗艦鍥婃場,鎻愬彇鍚庣殑寰場嫻撳害綰︿負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96.0%,CD44琛ㄨ揪鐜囦負50.2%,鑰孒LA-DR,CD34,CD29,CD73絳夎〃杈懼潎涓洪槾鎬，

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