人胚胎干细胞高效分化为巨核细胞模型的建立  

Establishment of a Highly Efficient Megakaryocytes Differentiation Model from Human Embryonic Stem Cells

作　　者：高洁 杨亦青[1,2] 庞素蕾 刘鑫 卢利莎[3] 雷晓华[4] 熊涛[3] 王洪涛 马峰[5] 周家喜 

机构地区：[1]中国医学科学院,北京协和医学院血液病医院(血液学研究所),实验血液学国家重点实验室,天津300020 [2]河北北方学院医学检验学院,张家口075000 [3]长江大学生命科学学院,荆州434025 [4]中国科学院动物研究所,北京100101 [5]中国医学科学院,北京协和医学院输血研究所,成都610052

出　　处：《中国细胞生物学学报》2014年第11期1470-1477,共8页Chinese Journal of Cell Biology

基　　金：天津市应用基础面上项目(批准号:11JCYBJC27300)资助的课题~~

摘　　要：人胚胎干细胞具有无限增殖潜能,且能够分化为包括巨核细胞在内的大多数的成熟血细胞。因此,为解决临床治疗血小板来源不足而建立一种由人胚胎干细胞高效分化获得成熟巨核细胞的分化模型具有重要应用前景。该文利用人胚胎干细胞与m AGM-S3基质细胞系共培养进行早期造血前体细胞的定向分化,并在分化第14 d获得大量"卵石样"造血前体细胞(CD43+细胞、CD45+细胞)。将"卵石样"造血前体细胞分离后,用无血清培养基分阶段添加SCF、TPO、IL-6、IL-3、FLt-3、IL-11等细胞因子进行巨核细胞定向诱导分化。结果显示,在分化的第3 d,CD41a+细胞比例可高达48.5%,CD42b+细胞比例可高达30.0%。对分化第6 d的细胞进行Wright-Giemsa染色,可见形态典型的巨核细胞。综上所述,该研究初步建立了一种人胚胎干细胞高效分化为巨核细胞的模型,为体外获得大量巨核细胞奠定了基础。Human embryonic stem cells（h ESCs） have unlimited proliferative capacity and are able to differentiate into all mature blood cell types including megakaryocytes.Hence,establishment of a highly efficient megakaryocytes differentiation model from hESCs may shed light on solving the problem of limited platelet source in clinical applications.Herein,we applied a model of h ESCs co-cultured with m AGM-S3 stromal cell line for hematopoietic differentiation.In this model,we could obtain abundant ＂cobblestone-like＂（CD43＋CD45＋cells） hematopoietic progenitor cells on the differentiation of day 14.Moreover,we isolated ＂cobblestone-like＂ hematopoietic progenitor cells and induced these cells to differentiate into megakaryocytes by adding cytokines,like SCF,TPO,IL-6,IL-3,FLt-3 and IL-11 by stages in serum free medium.Our research showed that the proportion of CD41a＋ cells could reach 48.5%,and the proportion of CD42b＋ cells could reach 30.0% on the differentiation of day 3.In addition,cells on the differentiation of day 6 showed characteristic of megakaryocytes by Wright-Giemsa staining.In conclusion,our research has preliminarily established a highly efficient megakaryocytes differentiation model from human embryonic stem cells,which provides a basis for obtaining plenty of megakaryocytes in vitro.

关 键 词：人胚胎干细胞 mAGM-S3 造血分化 巨核细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]