小鼠髓样抑制细胞体外扩增体系的建立  

Optimal conditions for the expansion of mouse myeloid-derived suppressor cells in vitro

作　　者：许秋桂 王文红[2] 朱波[1] 王瑞[1] 熊御云[1] 严佳婧 舒扬[1] 焦志军[1] 

机构地区：[1]江苏大学附属医院检验科,镇江212001 [2]江苏大学医学院病原生物学系,镇江212013

出　　处：《现代免疫学》2015年第2期140-144,共5页Current Immunology

基　　金：国家自然科学基金资助项目(81172871;81373217);江苏省科技厅科技创新与成果转化(生命健康科技)专项资金(BL2012057)

摘　　要：探讨体外诱导小鼠髓样抑制细胞(myeloid-derived suppressor cells,MDSC)扩增的方法及影响因素。采用不同浓度的细胞因子刺激不同浓度的新鲜骨髓细胞(1×105/ml、2.5×105/ml、5×105/ml),置于37℃、5%CO2培养不同时间(0、2、4d),流式细胞术检测MDSC的比例,并计数MDSC的绝对数。结果发现,三种浓度细胞因子组合刺激组MDSC的比例均显著高于新鲜骨髓组和单独GM-CSF或IL-6刺激组(P<0.05),但三组间未见明显差异;培养4d的MDSC比例亦明显高于培养2d的比例(P<0.05);骨髓细胞浓度为2.5×105/ml组诱导的MDSC比例高于其他两个细胞浓度组(P均<0.05)。本研究成功建立体外扩增MDSC的体系,为研究MDSC在肿瘤、炎症等疾病中的免疫机制奠定了基础。This research was designed to develop an effective method for the expansion of meloid-derived suppressor cells（MDSC）in vitro.（1,2.5,5）×105/ml fresh bone marrow cells were stimulated with various concentration combinations of IL-6and GM-CSF.Six concentration combinations were assigned which consisted of（GM-CSF（10ng/ml）＋ IL-6（10ng/ml）,GMCSF（20ng/ml）＋IL-6（20ng/ml）GM-CSF（40ng/ml）＋IL-6（40ng/ml）、GM-CSF（10ng/ml）,IL-6（10ng/ml））,cells were cultured for different times（0d,2d,4d）.The percentage of MDSC was determined using flow cytometry,and the absolute number of MDSC was also counted.The percentages of MDSC in cytokine treated groups were significantly higher than that of the fresh bone marrow group（P〈0.05）,however,there was no significant difference among three various concentrations of cytokine combination groups;The percentage of MDSC cultured for 4days was obviously higher compared with 2days（P〈0.05）;The concentrations of bone marrow cells had an effect on MDSC expansion,and the percentage of MDSC induced by2.5×105/ml bone marrow cells was significantly increased.This research has established an MDSC expansion system in vitro,which may be beneficial for further studying immune mechanisms of MDSC in diseases such as cancer or inflammation.

关 键 词：髓样抑制细胞 GM-CSF/IL-6 体外扩增 

分 类 号：R392.11[医药卫生—免疫学]