Induction of embryonic stem cells to hematopoietic cellsin vitro |
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Authors: | Ling Xu Chunping Qiao Shaoliang Huang Shunong Li Xuan Wu |
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Institution: | (1) Department of Pediatrics, Sun Yat-sen Memorial Hospital, Sun Yat-sen University of Medical Sciences, 510120 Guangzhou, China;(2) Department of Pathophysiology, Sun Yat-sen University of Medical Sciences, 510089 Guangzhou, China;(3) Department of Physiology, The Third Military Medical University, 400038 Chongqing, China |
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Abstract: | In order to get hematopoietic cells from embryonic stem (ES) cells and to study development mechanisms of hematopoietic cells,
the method of inducing embryonic stem cells to hematopoietic cells was explored by differenciating mouse ES cells and human
embryonic cells in three stages. The differentiated cells were identified by flow cytometry, immunohistochemistry and Wright’s
staining. The results showed that embryoid bodies (EBs) could form when ES cells were cultured in the medium with 2-mercaptoethanol
(2-ME). However, cytokines, such as stem cell factor (SCF), thrombopoietin (TPO), interleukin-3 (IL-3), interleukin-6 (IL-6),
erythropoietin (EPO) and granular colony stimulating factor (G-CSF), were not helpful for forming EBs. SCF, TPO and embryonic
cell conditional medium were useful for the differentiation of mouse EBs to hematopoietic progenitors. Eighty-six percent
of these cells were CD34+ after 6-d culture. Hematopoietic progenitors differentiated to B lymphocytes when they were cocultured with primary bone
marrow stroma cells in the DMEM medium with SCF and IL-6. 14 d later, most of the cells were CD34-CD38+. Wright’s staining and immunohistochemistry showed that 80% of these cells were plasma-like morphologically and immunoglubolin
positive. The study of hematopoietic cells from human embryonic cells showed that human embryonic cell differentiation was
very similar to that of mouse ES cells. They could form EBs in the first stage and the CD34 positive cells account for about
48.5% in the second stage. |
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Keywords: | embryonic stem cell cytokine hematopoietic stem cell |
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