Interleukin-3 promotes hemangioblast development in mouse aorta-gonad-mesonephros region
Interleukin-3 promotes hemangioblast development in mouse aorta-gonad-mesonephros region
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Interleukin-3促进小鼠主动脉-性腺-中肾区成血管细胞发育
DOI：
10.3324/haematol.2009.014241
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发表时间：
2010-06-01
期刊：
影响因子：
--
通讯作者：
Mao, Ning
中科院分区：
文献类型：
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作者：
He, Wen-Yan;Lan, Yu;Mao, Ning
背景成血管细胞是一种具有向造血细胞和血管细胞分化的双重潜能的前体细胞。在小鼠E7.0-7.5胚胎中，可以通过克隆母细胞集落形成细胞（BL-CFC）测定或单细胞0 P9共培养来鉴定成血管细胞。然而，在妊娠中期胚胎的成血管细胞的个体发育是不好defined.Design和MethodsThe BL-CFC检测和OP 9系统相结合，以说明成血管细胞与淋巴髓样和血管的潜力，在小鼠性腺-中肾区域。用集落形成试验、逆转录聚合酶链反应分析、免疫染色和流式细胞术鉴定其造血潜能，用基于Matrigel或OP 9的方法评价内皮祖细胞活性。结果在功能上，来源于性腺中肾的BL-CFC产生红系/髓系祖细胞、CD 19（+）B淋巴细胞和CD 3（+）TCR β（+）。T淋巴细胞。同时，BL-CFC来源的贴壁细胞在OP 9基质细胞上产生CD 31（+）管状结构，验证了内皮祖细胞的潜力。性腺-中肾来源的成血管细胞大量富集CD 31（+）、内粘蛋白（+）和CD 105（+）亚群，这些亚群共同确定内皮层为主要位置。有趣的是，在卵黄囊、胎盘、胎肝和胚胎循环中未检测到BL-CFC。筛选的候选细胞因子显示，白细胞介素3是显着的扩大BL-CFC的剂量依赖性的方式通过JAK 2/STAT 5和MAPK/ERK途径。中和白细胞介素-3在性腺-中肾区域导致BL-CFC的数量减少，表明这种细胞因子的生理需求。白细胞介素3处理的BL-CFC中造血和内皮分化潜能均显著增加，表明持续的积极影响。有趣的是，白细胞介素-3显着扩增原始红细胞和巨噬细胞前体在E7.5胚胎。定量聚合酶链反应分析表明下降Flk-1和升高Scl和von Willebrand因子转录后白细胞介素-3的刺激，表明加速hemangiopoies.ConclusionsThe hemangioblast与lymphomyeloid的潜力是在小鼠睾丸性腺中肾区域的造血的前体之一。白细胞介素-3对双能成血管细胞的数量和能力具有调节作用。
BackgroundThe hemangioblast is a bi-potential precursor cell with the capacity to differentiate into hematopoietic and vascular cells. In mouse E7.0-7.5 embryos, the hemangioblast can be identified by a clonal blast colony-forming cell (BL-CFC) assay or single cell OP9 co-culture. However, the ontogeny of the hemangioblast in mid-gestation embryos is poorly defined.Design and MethodsThe BL-CFC assay and the OP9 system were combined to illustrate the hemangioblast with lymphomyeloid and vascular potential in the mouse aorta-gonad-mesonephros region. The colony-forming assay, reverse transcriptase polymerase chain reaction analysis, immunostaining and flow cytometry were used to identify the hematopoietic potential, and Matrigel- or OP9-based methods were employed to evaluate endothelial progenitor activity.ResultsFunctionally, the aorta-gonad-mesonephros-derived BL-CFC produced erythroid/myeloid progenitors, CD19(+) B lymphocytes, and CD3(+)TCR beta(+). T lymphocytes. Meanwhile, the BL-CFC-derived adherent cells generated CD31(+) tube-like structures on OP9 stromal cells, validating the endothelial progenitor potential. The aorta-gonad-mesonephros-derived hemangioblast was greatly enriched in CD31(+), endomucin(+) and CD105(+) subpopulations, which collectively pinpoints the endothelial layer as the main location. Interestingly, the BL-CFC was not detected in yolk sac, placenta, fetal liver or embryonic circulation. Screening of candidate cytokines revealed that interleukin-3 was remarkable in expanding the BL-CFC in a dose-dependent manner through the JAK2/STAT5 and MAPK/ERK pathways. Neutralizing interleukin-3 in the aorta-gonad-mesonephros region resulted in reduced numbers of BL-CFC, indicating the physiological requirement for this cytokine. Both hematopoietic and endothelial differentiation potential were significantly increased in interleukin-3-treated BL-CFC, suggesting a persistent positive influence. Intriguingly, interleukin-3 markedly amplified primitive erythroid and macrophage precursors in E7.5 embryos. Quantitative polymerase chain reaction analysis demonstrated declined Flk-1 and elevated Scl and von Willebrand factor transcription upon interleukin-3 stimulation, indicating accelerated hemangiopoiesis.ConclusionsThe hemangioblast with lymphomyeloid potential is one of the precursors of definitive hematopoiesis in the mouse aorta-gonad-mesonephros region. Interleukin-3 has a regulatory role with regards to both the number and capacity of the dual-potential hemangioblast.