[Study on Wnt and Notch signalling involves in regulation of hematopoietic microenvironment.].
OBJECTIVE
To explore the mechanism of Wnt and Notch pathway involved modulating time and spatial restricted hematopoiesis.


METHODS
Murine hematopoietic stem and progenitor cells (HSPCs) were isolated from bone marrow (BM) by using c-kit microbeads. E10.5 aorta-gonad-mesonephros (AGM), E12.5, E14.5, E16.5 fetal liver (FL) and adult BM derived stromal cells (StroCs) were isolated and co-cultured with c-kit(+)HSPCs. The floating cells in co-culture system were sorted and counted by FACS. Gene expressions of Wnt and Notch pathway were detected by quantitative PCR and protein expressions by immunostaining.


RESULTS
Co-culturing HSPCs with AGM and FL-derived StroCs resulted in an expansion of c-kit(+)population from 0.4 x 10(5)/well to (19.2 +/- 3.2) x 10(5)/well and (26.8 +/- 5.4) x 10(5)/well, respectively, being greater than that with BM-derived StroCs (P < 0.05). The percentage of c-kit(+)cells detected in AGM- and BM- derived StroCs culture system was (75.2 +/- 7.1)%, (74.1 +/- 6.2)% respectively, being higher than FL- derived StroCs culture system (63.4 +/- 5.3)% (P < 0.05). Wnt and Notch pathway genes expression varied at different phases of hematopoiesis. Wnt was highly expressed in AGM and FL derived StroCs, and, Notch did in AGM and BM derived StroCs.


CONCLUSIONS
Wnt and Notch pathway are important modulators in regulating time and spatial restricted hematopoiesis.
Zhou
Kun
K
Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. Email: shanhum@163.com.
Hu
Cai-Hong
CH
Huang
Li-Fang
LF
Liu
Wen-Li
WL
Sun
Han-Ying
HY
D000818:Animals; D001011:Aorta / Q000166:cytology; D018920:Coculture Techniques; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D008650:Mesonephros / Q000166:cytology; D017154:Stromal Cells