[Bone marrow endothelial cell-conditioned medium promotes hematopoietic differentiation of mouse embryonic stem cells].
The purpose of this study was to observe the bone marrow endothelial cell-conditioned medium (BECM) and cytokines, i.e. vascular endothelial growth factor (VEGF), stem cell factor (SCF) and EPO promoting the generation of hematopoietic precursor cells from mouse embryonic stem cells (ESC) in vitro. Day 4 embryoid body (4dEB) cells were derived from ESC-D3 cell line, a murine ESC line, and then induced with BECM and/or cytokines. Four groups, i.e. BECM, BECM + VEGF + SCF + EPO, VEGF + SCF + EPO and control (spontaneous differentiation), were designed. Immunochemistry staining and flow cytometry were adopted to observe the antigen expression, RT-PCR to detect hematopoietic transcription factors, and hematopoietic progenitor assay to examine hematopoietic differentiation. The results showed that the cells induced from ESC expressed hematopoietic precursor cell antigens (c-kit, Sca-1, Thy-1 and CD34), transcription factors (c-myb, SCL and beta-H1) and generated HPP-CFC and BFU-E. The effect of BECM + VEGF + SCF + EPO was the most potent in the inducing groups according to the numbers of hematopoietic precursor cells and colonies. It is concluded that BECM promotes the differentiation of ESC into hematopoietic precursor cells in vitro, and this effect is the strongest when BECM combining with VEGF + SCF + EPO.
Zhao
Hui-Ping
HP
Laboratory of Blood Physiology, Xiangya Medical School of Centra South University, Changsha 410078, China.
Lu
Guang-Xiu
GX
Wang
Qi-Ru
QR
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D017077:Culture Media, Conditioned; D004622:Embryo, Mammalian / Q000166:cytology*; D042783:Endothelial Cells / Q000502:physiology*; D004921:Erythropoietin / Q000494:pharmacology; D005260:Female; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D051379:Mice; D019089:Stem Cell Factor / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology*; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology
D017077:Culture Media, Conditioned; D019089:Stem Cell Factor; D042461:Vascular Endothelial Growth Factor A; D004921:Erythropoietin