脐血间充质干细胞对PHA诱导的淋巴细胞增殖的影响

发布时间：2018-05-16 06:31

本文选题：脐血 + 骨髓　；参考：《南昌大学》2006年硕士论文

【摘要】：目的：了解脐血间充质干细胞(UCB-MSCs)对植物血凝素(PHA)诱导淋巴细胞(LCs)增殖的影响并与骨髓间充质干细胞(BM-MSCs)作比较；探讨UCB-MSCs对PHA诱导的LCs增殖抑制实验中的最佳细胞量效比；测定UCB-MSCs各亚组转化生长因子(TGF-β1)水平及肝素对UCB-MSCs分泌TGF-β1的影响。方法：对培养的UCB-MSCs和BM-MSCs进行形态、表型及功能方面的综合鉴定；将不同细胞浓度的UCB-MSCs和BM-MSCs加入由PHA诱导的LCs转化实验体系中，MTT法测定各亚组对LCs的抑制率。用ELISA法测定上清液中的TGF-β1水平。结果：(1)用密度梯度离心的方法从UCB和人BM中分离单个核细胞(MNC)，用贴壁法培养MSC，通过形态、表型及功能进行综合鉴定。培养至第3代的MSC具有长梭形的典型形态改变，表面分子检测显示高表达CD166、CD29、CD54，低表达CD13、CD34、CD45。经体外诱导可成为脂肪细胞。(2)MSCs：LCs为1：1、0.5：1、0.1：1、0.05：1、0.02：1、0.01：1时，UCB-MSCs组对PHA刺激的LCs增殖均有抑制作用，，抑制率分别为49.5±4.8、58.4±6.0、38.1±4.0、31.4±3.2、24.3±3.2、12.6±6.7；而BM-MSCs组抑制率随BM-MSCs数量不同而异，分别为52.4±8.4、65.1±9.7、34.7-±4.5、13.0±6.4、10.7±12.6、-43.9±9.4。UCB-MSCs组及BM-MSCs组在MSCs：LCs为0.05：1、0.02：1、0.01：1的三个亚组间对LCs的抑制率存在显著差异(P＜0.05)。UCB-MSCs组抑制率强于BM-MSCs组。(3)当UCB-MSCs：LCs为0.75：1时抑制作用最强，与其他亚组比较有显著差异(P＜0.05)。(4)UCB-MSCs各亚组上清液中TGF-β1水平高于BM-MSCs组，呈细胞剂量依赖性；UCB-MSCs组抑制率与TGF-β1水平相关性略小
[Abstract]:Objective: to investigate the effect of UCB-MSCs on the proliferation of lymphocytes in vitro induced by plant hemagglutinin (PHA) and compare it with that of BM-MSCs (BM-MSCs), and to explore the best dose-effectiveness ratio of UCB-MSCs in inhibiting the proliferation of LCs induced by PHA. The levels of TGF- 尾 1 in UCB-MSCs subgroups and the effect of heparin on TGF- 尾 1 secretion by UCB-MSCs were determined. Methods: the morphological phenotypic and functional characteristics of cultured UCB-MSCs and BM-MSCs were comprehensively identified and UCB-MSCs and BM-MSCs of different cell concentrations were added to the LCs transformation system induced by PHA to determine the inhibition rate of LCs in each subgroup. The level of TGF- 尾 1 in supernatant was determined by ELISA method. Results the mononuclear cells were isolated from UCB and human BM by density gradient centrifugation. The mononuclear cells were cultured by adherent method. The MSC cultured to the third generation had typical morphological changes of long spindle shape. Surface molecular analysis showed high expression of CD166T CD29 CD54 and low expression of CD13 CD34 + CD45. After induction in vitro, the MSCs: LCs were 1: 1: 0.5: 1: 0.1: 1: 1 0.05: 1. The inhibitory rates of UCB-MSCs on the proliferation of LCs stimulated by PHA were 49.5 卤4.88.4 卤6.038.1 卤4.038.1 卤4.038.1 卤4.038.1 卤3.21.The inhibitory rates of BM-MSCs group varied with the number of BM-MSCs. There were significant differences in the inhibition rate of LCs between the three subgroups with MSCs:LCs of 0.05: 1: 1: 0.02: 0.01: 1 (P < 0.05).UCB-MSCs) and BM-MSCs group (P < 3). The inhibition rate of LCs was significantly higher in 0.05).UCB-MSCs group than in BM-MSCs group (P < 0.05: 1) when UCB-MSCs:LCs was 0.75: 1. The inhibitory rate of LCs was significantly higher in the BM-MSCs group than in the BM-MSCs group (10.7 卤12.6 卤43.9 卤10.7 卤10.7 卤12.6ng-43.9 卤1.The MSCs:LCs was 0.05: 10.02: 1: 1, respectively, P < 0.05), but the inhibition rate in the 0.05).UCB-MSCs group was significantly higher than that in the BM-MSCs group. The level of TGF- 尾 1 in supernatant of each subgroup of 0.05).(4)UCB-MSCs was significantly higher than that of other subgroups, and the inhibition rate of UCB-MSCs was slightly correlated with the level of TGF- 尾 1 in a dose-dependent manner.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329.2

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