人胚胎干细胞高效诱导产生巨噬细胞方法的建立  

Establishment of an efficient method for large-scale production of mature macrophages from human embryonic stem cells

作　　者：郁金凤 孙文翠[1] 路旭琳 边国慧[1] 潘旭[1] 毛斌[1] 黄淑[1] 赖默温 周涯[1] 邹庆 李晴[1] 曾宪波[1] 李玉佳[1] 陈利民[1] 周家喜[3] 马峰[1,3] 

机构地区：[1]中国医学科学院北京协和医学院输血研究所,四川成都610052 [2]四川新生命干细胞科技股份有限公司 [3]中国医学科学院北京协和医学院血液学研究所实验血液学国家重点实验室

出　　处：《中国输血杂志》2015年第5期482-487,共6页Chinese Journal of Blood Transfusion

基　　金：国家自然科学基金(NSFC81170466);国家重点基础研究发展计划(973计划)资助项(2015CB964902)

摘　　要：目的利用人胚胎干细胞(h ESCs)建立高效巨噬细胞体外诱导分化体系,并与人脐带血CD34+细胞来源的巨噬细胞进行比较。方法通过将h ESCs与小鼠主动脉-性腺-中肾(m AGM)来源的基质细胞共培养产生CD34+CD45+造血干祖细胞,在多种细胞因子的诱导下悬浮培养14 d,利用May-Grünwald Giemsa染色、流式表面抗原及功能检测等方法对我们培养的巨噬细胞和人脐带血CD34+细胞来源的巨噬细胞进行比较鉴定。结果该方法可产生大量高纯度的巨噬细胞,成功的建立了体外高效诱导的巨噬细胞分化体系,并且该体系获得的巨噬细胞与人脐带血CD34+细胞来源的巨噬细胞具有相似的表型和功能。结论建立高效巨噬细胞体外诱导分化体系,为进一步研究人类巨噬细胞的早期发育和建立相关疾病模型提供了可靠的方法。Objective To establish an efficient co-culture system that can induce human embryonic stem cells （hESCs） to generate macmphages in vitro, to compare the hESC-derived macrophages with those derived from human cord blood CD34^＋ cells and to detect the morphology and function of the two-source maerophages. Methods An efficient method was proposed to generate mature macrophages by first co-culturing hESCs with mAGM-S3 （a murine AGM-derived stromal cell line） cells to produce CD34^＋ CD45 ^＋ multipotential hematopoietie progenitor cells. The suspension culture was then pooled in with various cytokines to differentiate myeloid cells for seven days and finally produced mature macrophages with M-CSF alone for another seven days. Moreover, these hESC-derived macrophages were compared with those derived from human cord blood CD34^＋ cells using May-Griinwald Giemsa staining, flow cytometry analysis and function evaluation experiments. Results Through this method, large quantity of macrophages could be produced （with a high purity -80% ）. Moreover, the study proved that these hESC-derived macrophages were normal in phenotype and function, similar to those derived from human cord blood CD34^＋ ceils. Conclusion Overall, the methods here established an efficient tool to conduct scalable experiments on human macrophages, which facilitates the genetic analysis on human macrophage-based disease model and the study on early developmental biology of human macrophages.

关 键 词：人胚胎干细胞(hESCs) 巨噬细胞 诱导分化 

分 类 号：R457.7[医药卫生—治疗学]