外源性bFGF和CNTF对大鼠MDSCs增殖的影响

发布时间：2018-03-29 13:44

  本文选题：MDSCs　切入点：bFGF　出处：《吉林大学》2009年硕士论文

【摘要】： 目前,MDSCs的研究存在许多热点问题,体外细胞培养中的纯化、扩增、定向分化等难题尚在探索中。人们在探讨适宜的干细胞培养条件的同时,特别关注生物活性因子对干细胞增殖的影响。bFGF和CNTF作为生物活性因子的成员,因其生物活性的多效性而备受基础与临床研究的重视。 本文进行的有关研究,旨在进一步探讨成体干细胞的培养条件,为有关研究与应用奠定实验基础。本实验通过酶消化法分离大鼠MDSCs,然后用密度梯度离心法和差速贴壁法纯化获取MDSCs,进行体外原代和传代培养。并通过免疫细胞化学法(desmin、Sca-1)对所获得的细胞进行鉴定。通过MTT比色法检测不同浓度(6.25、12.50、25.00、50.00、100.00ng/ml)的bFGF和CNTF单独应用96h以及二者(100.0ng/ml)联合作用24h、48h、72h和96h对MDSCs增殖的影响。MTT的OD值显示: 1.与阴性对照组比较,除6.25(ng/ml)组外,12.50ng/ml至100.00ng/ml实验组的MDSCs均有明显的增殖(P0.05)。在6.25ng/ml至50.00ng/ml期间,MDSCs的增殖随bFGF和CNTF浓度升高而显著增强(P0.05)。 2.在6.25ng/ml至50.00ng/ml期间,bFGF与CNTF对MDSCs的促增殖作用没有明显差异(P0.05)。随其浓度升高,在50.00ng/ml时,bFGF对MDSCs的促增殖作用明显低于CNTF组,并存在显著性差异(P0.05)。但50.00ng/ml组促增殖作用接近顶峰,与100.00ng/ml组比较,差异无统计学意义(P0.05)。 3.bFGF和CNTF的促增殖效应随培养时间的延长而逐渐明显,但促增殖作用的显效时间有明显差异。与阴性对照组比较,出现显著促增殖效应的时间(P0.05)分别为:bFGF单独作用96 h、CNTF单独作用72h、二者联合应用48h。实验组在24h内尽管MDSCs数量增多,但没有显著性差异(P0.05)。 4.在48h-96h期间,bFGF和CNTF联合组促进增殖的作用最强,并与其它实验组间存在显著差异(P0.05)。 本实验提示:bFGF和CNTF均可促进体外培养的大鼠MDSCs增殖,而且具有协同增效作用。但是,bFGF和CNTF促进MDSCs增殖作用具有显效时间的差异性,而且在12.5-50ng/ml期间,促MDSCs增殖作用还呈浓度依赖性。
[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.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：D919

【参考文献】

相关期刊论文 前10条

1 张晨晖;朱道立;;大鼠骨骼肌卫星细胞培养的研究[J];安徽农业科学;2008年12期

2 叶锦;靳风烁;陈锦;梁培禾;张克勤;;成年大鼠肌源性干细胞的培养和鉴定[J];第三军医大学学报;2006年10期

3 王特为,王廷华,刘芬,吴林艳;睫状神经营养因子的研究进展[J];中国组织化学与细胞化学杂志;2002年04期

4 吕萍;马景涛;;骨骼肌卫星细胞的分离及培养[J];河北医科大学学报;2006年04期

5 马焰,沈定国,赵民,江波,柳川;碱性成纤维生长因子和神经生长因子对骨骼肌成肌细胞增殖作用的研究[J];解放军医学杂志;1999年03期

6 张力;范明;王伟;陈晓萍;刘淑红;孙亮;;改良法体外培养大鼠成肌细胞的实验研究[J];军事医学科学院院刊;2007年01期

7 刘福强;刘畅;杨彪;梅晰凡;;新生大鼠肌源性干细胞的原代培养和鉴定[J];辽宁医学院学报;2008年01期

8 郭雨霁,李盛芳;神经营养因子家族及其受体的研究进展[J];神经解剖学杂志;2001年03期

9 许家军,陈尔瑜,路长林,姜宗来,何成;重组睫状神经营养因子对周围神经再生中多种细胞的作用[J];神经解剖学杂志;2002年04期

10 何成，路长林，敖世洲;睫状神经营养因子研究进展[J];生理科学进展;1996年01期

，

本文编号：1681382