本文選題：MDSCs　切入點(diǎn)：bFGF　出處：《吉林大學(xué)》2009年碩士論文

【摘要】： 目前,MDSCs的研究存在許多熱點(diǎn)問(wèn)題,體外細(xì)胞培養(yǎng)中的純化、擴(kuò)增、定向分化等難題尚在探索中。人們?cè)谔接戇m宜的干細(xì)胞培養(yǎng)條件的同時(shí),特別關(guān)注生物活性因子對(duì)干細(xì)胞增殖的影響。bFGF和CNTF作為生物活性因子的成員,因其生物活性的多效性而備受基礎(chǔ)與臨床研究的重視。 本文進(jìn)行的有關(guān)研究,旨在進(jìn)一步探討成體干細(xì)胞的培養(yǎng)條件,為有關(guān)研究與應(yīng)用奠定實(shí)驗(yàn)基礎(chǔ)。本實(shí)驗(yàn)通過(guò)酶消化法分離大鼠MDSCs,然后用密度梯度離心法和差速貼壁法純化獲取MDSCs,進(jìn)行體外原代和傳代培養(yǎng)。并通過(guò)免疫細(xì)胞化學(xué)法(desmin、Sca-1)對(duì)所獲得的細(xì)胞進(jìn)行鑒定。通過(guò)MTT比色法檢測(cè)不同濃度(6.25、12.50、25.00、50.00、100.00ng/ml)的bFGF和CNTF單獨(dú)應(yīng)用96h以及二者(100.0ng/ml)聯(lián)合作用24h、48h、72h和96h對(duì)MDSCs增殖的影響。MTT的OD值顯示: 1.與陰性對(duì)照組比較,除6.25(ng/ml)組外,12.50ng/ml至100.00ng/ml實(shí)驗(yàn)組的MDSCs均有明顯的增殖(P0.05)。在6.25ng/ml至50.00ng/ml期間,MDSCs的增殖隨bFGF和CNTF濃度升高而顯著增強(qiáng)(P0.05)。 2.在6.25ng/ml至50.00ng/ml期間,bFGF與CNTF對(duì)MDSCs的促增殖作用沒(méi)有明顯差異(P0.05)。隨其濃度升高,在50.00ng/ml時(shí),bFGF對(duì)MDSCs的促增殖作用明顯低于CNTF組,并存在顯著性差異(P0.05)。但50.00ng/ml組促增殖作用接近頂峰,與100.00ng/ml組比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。 3.bFGF和CNTF的促增殖效應(yīng)隨培養(yǎng)時(shí)間的延長(zhǎng)而逐漸明顯,但促增殖作用的顯效時(shí)間有明顯差異。與陰性對(duì)照組比較,出現(xiàn)顯著促增殖效應(yīng)的時(shí)間(P0.05)分別為:bFGF單獨(dú)作用96 h、CNTF單獨(dú)作用72h、二者聯(lián)合應(yīng)用48h。實(shí)驗(yàn)組在24h內(nèi)盡管MDSCs數(shù)量增多,但沒(méi)有顯著性差異(P0.05)。 4.在48h-96h期間,bFGF和CNTF聯(lián)合組促進(jìn)增殖的作用最強(qiáng),并與其它實(shí)驗(yàn)組間存在顯著差異(P0.05)。 本實(shí)驗(yàn)提示:bFGF和CNTF均可促進(jìn)體外培養(yǎng)的大鼠MDSCs增殖,而且具有協(xié)同增效作用。但是,bFGF和CNTF促進(jìn)MDSCs增殖作用具有顯效時(shí)間的差異性,而且在12.5-50ng/ml期間,促M(fèi)DSCs增殖作用還呈濃度依賴(lài)性。
[Abstract]:At present, there are many hot problems in the research of MDSCs, such as purification, amplification, directional differentiation and so on. The effects of bioactive factors on the proliferation of stem cells. BFGF and CNTF as members of bioactive factors have attracted much attention in basic and clinical studies because of their biological activity. The purpose of this study is to further explore the culture conditions of adult stem cells. In this experiment, MDSCs were isolated by enzyme digestion, then purified by density gradient centrifugation and differential adhesion method. MTT colorimetric assay was used to detect the effects of bFGF and CNTF at different concentrations on the proliferation of MDSCs for 96 h and 24 h, 48 h, 72 h and 96 h, respectively. 1.Compared with the negative control group, the proliferation of MDSCs in the 100.00ng/ml group was significantly increased with the increase of bFGF and CNTF concentrations, except for the 6.25ng / ml group, and the proliferation of P0.05G / ml in the 100.00ng/ml group was significantly increased with the increase of bFGF and CNTF concentration during the period of 6.25ng/ml to 50.00ng/ml. 2. There was no significant difference between CNTF and 6.25ng/ml in promoting the proliferation of MDSCs between 6.25ng/ml and 50.00ng/ml. With the increase of 6.25ng/ml concentration, the proliferation of MDSCs in 50.00ng/ml group was significantly lower than that in CNTF group, and there was significant difference (P 0.05). However, the proliferation effect of 50.00ng/ml group was near the peak. There was no significant difference compared with 100.00ng/ml group (P 0.05). The proliferative effect of 3.bFGF and CNTF was more and more obvious with the increase of culture time, but there was significant difference between the two groups. The time of significant proliferative effect (P0.05) was 96 h and 72 h, respectively. In the experimental group, the number of MDSCs increased, but there was no significant difference in the number of MDSCs in the experimental group (P 0.05). 4. During 48h-96h, the combination of bFGF and CNTF had the strongest effect on promoting proliferation, and there was a significant difference between the two groups and other experimental groups (P 0.05). The results suggest that both CNTF and BFGF can promote the proliferation of MDSCs in vitro and have synergistic effect. However, the effects of bFGF and CNTF on the proliferation of MDSCs have significant difference in time, and during the period of 12.5-50ng/ml, the effects of BFGF and CNTF on the proliferation of MDSCs were significantly different. The proliferative effect of MDSCs was also concentration-dependent.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類(lèi)號(hào)】：D919

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張晨暉;朱道立;;大鼠骨骼肌衛(wèi)星細(xì)胞培養(yǎng)的研究[J];安徽農(nóng)業(yè)科學(xué);2008年12期

2 葉錦;靳風(fēng)爍;陳錦;梁培禾;張克勤;;成年大鼠肌源性干細(xì)胞的培養(yǎng)和鑒定[J];第三軍醫(yī)大學(xué)學(xué)報(bào);2006年10期

3 王特為,王廷華,劉芬,吳林艷;睫狀神經(jīng)營(yíng)養(yǎng)因子的研究進(jìn)展[J];中國(guó)組織化學(xué)與細(xì)胞化學(xué)雜志;2002年04期

4 呂萍;馬景濤;;骨骼肌衛(wèi)星細(xì)胞的分離及培養(yǎng)[J];河北醫(yī)科大學(xué)學(xué)報(bào);2006年04期

5 馬焰,沈定國(guó),趙民,江波,柳川;堿性成纖維生長(zhǎng)因子和神經(jīng)生長(zhǎng)因子對(duì)骨骼肌成肌細(xì)胞增殖作用的研究[J];解放軍醫(yī)學(xué)雜志;1999年03期

6 張力;范明;王偉;陳曉萍;劉淑紅;孫亮;;改良法體外培養(yǎng)大鼠成肌細(xì)胞的實(shí)驗(yàn)研究[J];軍事醫(yī)學(xué)科學(xué)院院刊;2007年01期

7 劉福強(qiáng);劉暢;楊彪;梅晰凡;;新生大鼠肌源性干細(xì)胞的原代培養(yǎng)和鑒定[J];遼寧醫(yī)學(xué)院學(xué)報(bào);2008年01期

8 郭雨霽,李盛芳;神經(jīng)營(yíng)養(yǎng)因子家族及其受體的研究進(jìn)展[J];神經(jīng)解剖學(xué)雜志;2001年03期

9 許家軍,陳爾瑜,路長(zhǎng)林,姜宗來(lái),何成;重組睫狀神經(jīng)營(yíng)養(yǎng)因子對(duì)周?chē)窠?jīng)再生中多種細(xì)胞的作用[J];神經(jīng)解剖學(xué)雜志;2002年04期

10 何成，路長(zhǎng)林，敖世洲;睫狀神經(jīng)營(yíng)養(yǎng)因子研究進(jìn)展[J];生理科學(xué)進(jìn)展;1996年01期

，

本文編號(hào)：1681382