ERK信号通路在人胚胎干细胞造血分化中的作用研究  

The Role of ERK Signaling in Hematopoietic Differentiation from hESCs

作　　者：王洪涛[1] 刘鑫[1] 王梦鸽[1] 苏培[1] 张磊升[1] 刘翠翠[1] 王钰[1] 吴丹[1] 涂茜[1] 周家喜[1] Wang Hongtao Liu Xin Wang Mengge Su Pei Zhang Leisheng Liu Cuicui Wang Yu Wu Dan Tu Qian Zhou Jiaxi(Stata Key Laboratory of Experimental Hematology, Institute of Hematology ＆ Blood Diseases Hospital Chinese Academy of Medical Sciences ＆ Peking Union Medical College, Tianjin 300020, Chin)

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

出　　处：《中国细胞生物学学报》2017年第6期725-731,共7页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:31500949);中国医学科学院医学与健康科技创新工程项目(批准号:2016-I2M-1-018,2016-I2M-3-002)资助的课题~~

摘　　要：细胞外调节蛋白激酶(extracellular regulated protein kinase,ERK)信号通路在斑马鱼及小鼠造血发生中发挥着重要作用,但其在人胚胎干细胞(human embryonic stem cells,h ESCs)造血分化中的作用尚不清楚。该研究利用h ESCs单层造血分化模型及ERK信号通路抑制剂PD98059探索了ERK信号通路在h ESCs造血分化中的作用。采用免疫荧光技术、流式细胞术分析发现,PD98059能够显著抑制CD43^+造血干/祖细胞的产生。进一步的研究发现,PD98059的作用阶段为APLNR+侧板中胚层产生阶段,在该作用阶段添加PD98059与造血分化全程添加对CD43^+造血干/祖细胞产生的抑制效果一致。该研究结果表明,抑制ERK信号通路通过抑制侧板中胚层细胞的产生而抑制h ESCs造血分化。该研究为建立体外h ESCs高效造血分化体系及规模化产生功能性血细胞奠定了理论基础。Extracellular regulated protein kinase（ERK） pathway has been reported to play a critical role in hematopoiesis in zebrafish and mouse. However, the role of ERK pathway in hematopoietic differentiation from human embryonic stem cells（h ESCs） remains unclear. In this study, we investigated the role of ERK pathway in hematopoietic specification from h ESCs by taking the advantage of a chemical defined hematopoietic differentiation model and PD98059, a specific inhibitor of ERK pathway. The results of immunofluorescence and flow cytometry analysis revealed that PD98059 treatment significantly inhibited the generation of CD43 positive hematopoietic stem and progenitor cells（HSPCs）. Further studies demonstrate that PD98059 suppresses hematopoietic differentiation through inhibiting the generation of APLNR positive lateral plate mesoderm cells. The addition of PD98059 in the stage of lateral plate mesoderm cell generation and the whole process of hematopoietic differentiation shows similar inhibitory effect on the generation of HSPCs. Taken together, our findings indicate that the inhibition of ERK pathway suppresses hematopoietic differentiation from h ESCs through repressing the generation of lateral plate mesoderm cells. This study provides a theoretical foundation for the establishment of a highly efficient hematopoietic differentiation system of h ESCs and the large-scale production of functional blood cells in vitro.

关 键 词：人胚胎干细胞 造血分化 ERK信号通路 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]