[Microenvironment for the hematopoietic differentiation of human embryonic stem cells].
OBJECTIVE
To investigate the roles of human fetal liver stromal cells (hFLSCs) and human fetal bone marrow stromal cells (hFBMSCs) in the hematopoietic differentiation of human embryonic stem cells and analyze their gene expression profile changes.


METHODS
The embryonic bodies on day 4 were cocultured with hFLSCs or hFBMSC in the presence of cytokines. Flow cytometry was performed after 8 days of induction to detect the expressions of the hemangioblast markers KDR and CD34, and the differential gene expression profiles between hFBMSC and hFLSCs were examined by cDNA microarray analysis.


RESULTS
Eight days after the induction, (1.06-/+0.20)% of the hFLSCs and (8.8-/+1.49)% of the hFBMSCs were positive for KDR, with the positivity rates for CD34 of (1.25-/+0.16)% and (9.17-/+2.10)%, respectively. In hFLSCs and hFBMSCs cultures, 0.9-/+0.36 and 10.6-/+0.63 hemagioblast-like cell colonies were found, respectively. cDNA microarray analysis showed that 240 genes were highly expressed in hFBMSCs, and 21 genes related to secreted cytokines, cell adhesion molecules and extracellular matrix proteins were highly expressed.


CONCLUSIONS
The microenvironment including the cell matrix protein and cytokines secreted by the hFBMSCs might play an important role in hemangioblastic differentiation of human bone marrow stromal cells in vitro.
Wang
Jian
J
Institute of Reproduction and Stem Cell Engineering, Central South University, State Human Stem Cell Engineering Research Center, Changsha, China. wjlx2000@yahoo.com.cn
Ling
Ge
G
Zhao
Hui-Ping
HP
Lu
Guang-Xiu
GX
D018952:Antigens, CD34 / Q000235:genetics / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation / Q000502:physiology*; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology*; D005333:Fetus; D020869:Gene Expression Profiling; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D008099:Liver / Q000166:cytology*; D020411:Oligonucleotide Array Sequence Analysis; D017154:Stromal Cells / Q000502:physiology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism
D018952:Antigens, CD34; D040301:Vascular Endothelial Growth Factor Receptor-2