發(fā)布時(shí)間：2018-02-24 14:19

  本文關(guān)鍵詞： 血小板衍生膜微粒 臍帶血 粒-巨噬祖細(xì)胞集落 增殖　出處：《東南大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的研究血小板衍生膜微粒(PMPs)的制備方法,并探討PMPs對(duì)人臍帶血造血干細(xì)胞增殖能力的影響。 方法分別采用不同濃度的凝血酶激活血小板釋放出PMPs,采用流式細(xì)胞儀檢測(cè)PMPs的釋放量,確定凝血酶最佳實(shí)驗(yàn)濃度。應(yīng)用掃描電鏡觀察凝血酶激活血小板后PMPs的釋放情況。采用Ficoll分離人臍帶血單個(gè)核細(xì)胞(MNCs),在含有不同濃度PMPs或血小板的甲基纖維素半固體培養(yǎng)基中培養(yǎng)7天,觀察人臍帶血粒-巨噬祖細(xì)胞集落(CFU-GM)形成情況,并用流式細(xì)胞儀檢測(cè)在含PMPs的無(wú)血清培養(yǎng)體系中培養(yǎng)7天后人臍帶血MNCs的CD34抗原表達(dá)的情況。 結(jié)結(jié)果用2.0 U/ml、1.5 U/ml、1.0 U/ml和0.5 U/ml凝血酶激活血小板后的PMPs釋放率分別為:28.7%、47.7%、50.1%和43.9%;CFU-GM產(chǎn)率與PMPs呈劑量依賴關(guān)系,PMPs為10μg/ml、50μg/ml、100μg/ml和血小板為200×109/L時(shí)的CFU-GM產(chǎn)率(個(gè)/2×105 MNCs)分別為:119.8±32.2、142.9±45.2、180.8±85.1和136.5±43.6, 50μg/ml、100μg/ml PMPs和200×109/L血小板組的集落產(chǎn)率與空白對(duì)照組(103.0±24.8)相比,P值均0.05;而10μg/ml PMPs組的集落產(chǎn)率略高于空白對(duì)照組,但兩者無(wú)統(tǒng)計(jì)學(xué)差異,P值0.05。無(wú)血清培養(yǎng)7天后,50μg/ml PMPs組CD34抗原表達(dá)率為:0.73%±0.74%,與空白對(duì)照組(0.43%±0.54%)相比,P值0.05。 結(jié)論用1.0 U/ml的凝血酶激活血小板釋放的PMPs較為均一且釋放量最大。PMPs可促進(jìn)人臍帶血粒-巨噬祖細(xì)胞的增殖和CD34抗原的表達(dá)。
[Abstract]:Objective to study the preparation of platelet-derived membrane (PMPs) and the effect of PMPs on the proliferation of human umbilical cord blood hematopoietic stem cells. Methods different concentrations of thrombin were used to activate platelet to release PMPs. Flow cytometry was used to detect the release of PMPs. The optimal concentration of thrombin was determined. The release of PMPs after thrombin activation was observed by scanning electron microscope. Ficoll was used to isolate mononuclear cells from human umbilical cord blood and methylcellulose hemicellulose containing different concentrations of PMPs or platelets was obtained. Culture in solid medium for 7 days, The formation of human umbilical cord blood granulocyte-macrophage colony (CFU-GM) was observed, and the expression of CD34 antigen of MNCs was detected by flow cytometry in serum-free culture system containing PMPs for 7 days. The PMPs release rates were 47.70.1% and 43.9% in a dose-dependent relationship with 10 渭 g / ml 50 渭 g / ml 50 渭 g / ml 50 渭 g / ml CFU-GM and 200 脳 10 ~ (5) MNCs, respectively. The PMPs release rates were: 119.8 鹵32.2142.9 鹵45.2180.8 鹵85.1 and 136.5 鹵43.6ml, 50 渭 g / ml, 100 渭 g / ml PMPs and 50 渭 g / ml / ml, respectively119.8 鹵32.2142.9 鹵45.2180.8 鹵85.1 and 136.5 鹵43.6ml / ml, 50 渭 g / ml, 100 渭 g / ml PMPs and 50 渭 g / ml / ml / ml of platelets / platelets, 200 脳 109Mncml / ml and 200 脳 109Mncml / ml of platelets, respectively519.8 鹵32.2142.9 鹵45.2142.9 鹵45.2180.8 鹵85.1 and 136.5 鹵43.60.50 渭 g / ml respectively. Compared with the control group (103.0 鹵24.8), the colony production rate of the 200 脳 10 9 / L platelet group was 0.05, while the colony yield rate of the 10 渭 g / ml PMPs group was slightly higher than that of the blank control group, and that of the 10 渭 g / ml PMPs group was slightly higher than that of the blank control group. After 7 days of serum-free culture, the expression rate of CD34 antigen in 50 渭 g / ml PMPs group was 0.73% 鹵0.74%, which was higher than that in the blank control group (0.43% 鹵0.54%). Conclusion PMPs released from platelets activated by 1.0 U / ml thrombin can promote the proliferation of human umbilical cord blood granulocyte-macrophage and the expression of CD34 antigen.
【學(xué)位授予單位】：東南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R329.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 孟二紅,吳祖澤,王立生;膜微粒子與造血調(diào)控[J];中國(guó)實(shí)驗(yàn)血液學(xué)雜志;2005年04期

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本文編號(hào)：1530554