无细胞接触条件下成人骨髓间充质干细胞培养上清对异体淋巴细胞增殖的负调控作用(英文)  

Negatively regulatory effects of bone marrow-derived mesenchymal stem cells supernatant on allogeneic lymphocytes proliferation

作　　者：吴涛[1] 石静云[2] 白海[1] 王存邦[1] 路继红[1] 欧剑锋[1] 

机构地区：[1]解放军兰州军区兰州总医院全军血液病中心,甘肃省兰州市730050 [2]甘肃省妇幼保健院新生儿重症救护中心,甘肃省兰州市730050

出　　处：《中国组织工程研究与临床康复》2008年第43期8579-8582,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：Supported by: grants from the Military the-Eleventh-Five- Year Project Elite Foundation, No. 06J005;grants from the Military the-Eleventh-Five-year Project Important Foundation, No. 06MB099~~

摘　　要：背景:成人骨髓间充质干细胞对异体淋巴细胞增殖具有负调控作用的理论已公认,但证实其具体机制的资料有限。目的:在无细胞接触的条件下,观察成人骨髓间充质干细胞培养上清对异体淋巴细胞增殖的负调控效果。设计、时间及地点:细胞学体外观察,于2005-10/2007-12在解放军兰州军区兰州总医院完成。材料:骨髓标本来源于行异基因造血干细胞移植的健康供者,外周血来源于健康志愿者。方法:无菌条件下采集成人骨髓,Percoll+贴壁法体外分离培养骨髓间充质干细胞。取新鲜肝素抗凝的外周血,Ficoll法分离出淋巴单个核细胞,调整浓度为2×109L-1,分别取100μL淋巴细胞悬液,置入96孔板中,设立4组:培养上清组加入培养3d的骨髓间充质干细胞培养上清,100μL/孔;培养上清+植物血凝素组在前组基础上加入1g/L植物血凝素5μL;空白对照组加入含体积分数0.1胎牛血清的LG-DMEM培养液100μL;植物血凝素组在空白对照组基础上加入1g/L植物血凝素5μL。置37℃、体积分数为0.05的CO2饱和湿度孵箱中培养3d。主要观察指标:骨髓间充质干细胞培养上清对异体淋巴细胞增殖转化的影响。结果:与空白对照组和植物血凝素组比较,加入骨髓间充质干细胞培养上清后淋巴细胞增殖活性明显下降(P<0.05),其增殖转化抑制率为9.00%。与培养上清组比较,在有植物血凝素刺激的情况下,淋巴细胞增殖活性进一步下降(P<0.01),其增殖转化抑制率达20.91%。结论:成人骨髓间充质干细胞培养上清能够抑制异体淋巴细胞增殖转化,且在外源性刺激源植物血凝素存在的情况下,其抑制效果明显增强,提示其负调控作用至少是部分通过分泌可溶性细胞因子而间接对淋巴细胞产生抑制的。BACKGROUND： Preliminary study has proven that adult human bone marrow-derived mesenchymal stem cells （MSCs） suppress peripheral blood lymphocytes proliferation. But the mechanism was still to be investigated. OBJECTIVE： To study the negatively regulatory effect of adult human MSCs on allogeneic lymphocyte proliferation by cell-free condition. DESIGN, TIME AND SETTING： Cytological observation in vitro, which was performed in the Lanzhou General Hospital, Lanzhou Military Area Command of Chinese PLA between October 2005 and December 2007. MATERIALS： The bone marrow sample was provided by the allo-transplantation donor. The peripheral blood lymphocytes were provided by the healthy volunteer. METHODS： Adult human MSCs were separated with Percoll ＋ adherence method. Allogeneic peripheral blood lymphocytes were obtained from healthy donors with Ficoll solution and the cell concentration was adjustedas 2×10^9/L for use. 100 μL MSCs culture supernatant was taken out in 96-well plates. The groups were following： A supernatant ＋ 3-day MSCs culture media （100 μL/well）; B supernatant ＋ phytohemagglutini （PHA; 1 g/L, 5 μL）; C medium ＋ LG-DMEM culture media containing 10% fetal bovine serum （100 μL）; D medium ＋ PHA （1 g/L, 5 μL）. The cells were incubated at 37 ℃ with 5% CO2 in a fully humidified atmosphere for three days. MAIN OUTCOME MEASURES： Effect of MSCs supernatant on proliferation and transformation of variant lymphocytes. RESULTS： Peripheral blood lymphocytes proliferation was suppressed as compared with the blank control group and PHA group after MSCs culture, and the inhibition ratio was 9.00% （P 〈 0.05）. When lymphocytes were stimulated by PHA, the suppression effects were even stronger and the inhibition ratio was 20.91% （P 〈 0.01）. CONCLUSION ： Adult human MSCs supernatant can suppress peripheral blood lymphocytes proliferation and transformation; furthermore, pHA can enhance the inhibitory effect, suggesting the negative regulation is at least in

关 键 词：骨髓间充质干细胞 免疫调控 淋巴细胞 

分 类 号：R394.2[医药卫生—医学遗传学]