Zhou Kun, Hu Cai-Hong, Huang Li-Fang, Liu Wen-Li, Sun Han-Ying
Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. Email:
Zhonghua Xue Ye Xue Za Zhi. 2009 Dec;30(12):799-803.
To explore the mechanism of Wnt and Notch pathway involved modulating time and spatial restricted hematopoiesis.
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.
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.
Wnt and Notch pathway are important modulators in regulating time and spatial restricted hematopoiesis.
探讨Wnt和Notch信号通路参与调控造血时空特异性的机制。
采用c-kit微珠从骨髓中分离小鼠造血干细胞和祖细胞（HSPCs）。分离E10.5期主动脉-性腺-中肾（AGM）、E12.5、E14.5、E16.5期胎肝（FL）及成年骨髓来源的基质细胞（StroCs），并与c-kit(+)HSPCs共培养。通过流式细胞术对共培养体系中的悬浮细胞进行分选和计数。采用定量PCR检测Wnt和Notch信号通路的基因表达，免疫染色检测蛋白表达。
HSPCs与AGM和FL来源的StroCs共培养后，c-kit(+)细胞群分别从0.4×10⁵/孔扩增至(19.2±3.2)×10⁵/孔和(26.8±5.4)×10⁵/孔，均高于与BM来源的StroCs共培养组（P<0.05）。AGM和BM来源的StroCs培养体系中检测到的c-kit(+)细胞百分比分别为(75.2±7.1)%、(74.1±6.2)%，均高于FL来源的StroCs培养体系（63.4±5.3%）（P<0.05）。Wnt和Notch信号通路基因在造血的不同阶段表达各异。Wnt在AGM和FL来源的StroCs中高表达；Notch在AGM和BM来源的StroCs中高表达。
Wnt和Notch信号通路是调控造血时空特异性的重要调节因子。
