Home > Browse Issues > Vol.36 No.11
Establishment of a Highly Efficient Megakaryocytes Differentiation Model from Human Embryonic Stem Cells
Gao Jie1#, Yang Yiqing1,2#, Pang Sulei1, Liu Xin1, Lu Lisha3, Lei Xiaohua4, Xiong Tao3, Wang Hongtao1,Ma Feng5, Zhou Jiaxi1*
1State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China;
2Hebei North University, Faculty of Medical Laboratory Sciences,ZhangJiaKou 075000, China;
3College of Life Sciences, Yangtze University, Jingzhou 434025,China;
4Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China;
5Institute of Blood Transfusion, Chinese Academy of Medical Sciences & Peking Union Medical College, Chengdu 610052, China
Abstract: Human embryonic stem cells (hESCs) 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 sourcein clinical applications. Herein， we applied a model of hESCs co-cultured with mAGM-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.
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.
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