[BMP-4 and VEGF promote development of hematopoietic stem cells during the embryoid body formation from embryonic stem cells].
The objective of this study was to explore the effects of BMP-4 and VEGF on the development of primary hematopoietic stem cells during the differentiation of embryonic stem cells (ESCs) into embryoid body (EB). Murine E14 ESCs were seeded into semisolid methylcellulose-based medium for EB formation. According to added or not cytokines, experiments were divided into: (1) group of spontaneous differentiation without cytokine as control; (2) group of BMP-4 in different concentrations (0, 5, 15, 25 and 50 ng/ml); (3) group of BMP-4 combined with VEGF; (4) group of VEGF alone. EBs were collected on days 3, 6, 9, 12, 15, and the proportion of Flk-1(+) cells were assayed by flow cytometry. The results showed that in the different BMP-4 concentration groups, the proportions of Flk-1(+) cells were significantly different, and it reached the peak values in 25 ng/ml BMP-4 group as 6.51 +/- 1.02% at day 3 and 7.70 +/- 1.12% at day 6 respectively, which were statistically higher than those in control group without-BMP-4 and in 5 ng/ml BMP-4 group (p < 0.05). When BMP-4 was used in combination with VEGF, Flk-1(+) cells went to peak proportion value at day 9 as 27.53 +/- 8.14%, which was statistically higher than that in spontaneous differentiation group as 8.77 +/- 2.35% (p < 0.05) and VEGF treatment group as 11.21 +/- 2.23% (p < 0.05). It is concluded that BMP-4 in combination with VEGF can promote Flk-1(+) cells genesis during EB formation in vitro, which provides experimental evidence for researches on directed differentiation of ESCs into hematopoietic stem cells simulating the microenvironment in vivo.
Chen
Hui-Qin
HQ
Department of Pediatrics, The Third Hospital, SUN Yat-sen University, Guangzhou 510630, Guangdong Province, China.
Zhang
Xu-Chao
XC
Huang
Shao-Liang
SL
Cai
Yun
Y
Wu
Bei-Yan
BY
Zhou
Dun-Hua
DH
Huang
Ke
K
D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D051379:Mice; D042442:Vascular Endothelial Growth Factors / Q000494:pharmacology*
C525810:Bmp4 protein, mouse; D055415:Bone Morphogenetic Protein 4; D042442:Vascular Endothelial Growth Factors