Vascular niche promotes hematopoietic multipotent progenitor formation from pluripotent stem cells
Vascular niche promotes hematopoietic multipotent progenitor formation from pluripotent stem cells
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DOI：
10.1172/jci79328
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发表时间：
2015-03-01
影响因子：
15.9
通讯作者：
Kiem, Hans-Peter
中科院分区：
文献类型：
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作者：
Gori, Jennifer L.;Butler, Jason M.;Kiem, Hans-Peter
多能干细胞（PSCs）是治疗造血疾病的一种替代造血干细胞（HSC）来源。人类psc衍生的多能祖细胞（MPP）的有限移植阻碍了这些细胞的临床应用，并表明MPP需要额外的线索来确定造血。我们假设血管生态位的存在能够产生Notch配体jaggepped -1 （JAG1）和delta-like配体4 (DLL4)，从而驱动最终的造血。我们用表达JAG1和DLL4的内皮细胞（ECs）存在或不存在的细胞因子分化hes2人胚胎干细胞（hESC）和Macaca nemestrina诱导的PSC (iPSC) line-7。与不含ECs或缺乏JAG1或DLL4的ECs共培养的细胞相比，与ECs共培养的细胞产生了更多的CD34(+)CD45（+）造血祖细胞。ec诱导的细胞表现出Notch激活并表达hsc特异性Notch靶点RUNX1和GATA2。与细胞因子诱导的细胞相比，ec诱导的PSC-MPP在NOD/SCID/IL-2受体γ链缺失（NSG）小鼠中的植入水平明显更高，并且基于低剂量化疗的选择进一步增加了植入。通过血管生态位诱导实现的长期植入和髓淋巴比与脐带血来源的MPP所达到的水平相似，比hscs来源的MPP所达到的水平高出20倍。我们的研究结果表明，内皮Notch配体促进psc最终的造血和长期移植CD34（+）细胞的产生，表明这些配体对HSC的出现至关重要。
Pluripotent stem cells (PSCs) represent an alternative hematopoietic stem cell (HSC) source for treating hematopoietic disease. The limited engraftment of human PSC-derived (hPSC-derived) multipotent progenitor cells (MPP) has hampered the clinical application of these cells and suggests that MPP require additional cues for definitive hematopoiesis. We hypothesized that the presence of a vascular niche that produces Notch ligands jagged-1 (JAG1) and delta-like ligand-4 (DLL4) drives definitive hematopoiesis. We differentiated hes2 human embryonic stem cells (hESC) and Macaca nemestrina-induced PSC (iPSC) line-7 with cytokines in the presence or absence of endothelial cells (ECs) that express JAG1 and DLL4. Cells cocultured with ECs generated substantially more CD34(+)CD45(+) hematopoietic progenitors compared with cells cocultured without ECs or with ECs lacking JAG1 or DLL4. EC-induced cells exhibited Notch activation and expressed HSC-specific Notch targets RUNX1 and GATA2. EC-induced PSC-MPP engrafted at a markedly higher level in NOD/SCID/IL-2 receptor gamma chain-null (NSG) mice compared with cytokine-induced cells, and low-dose chemotherapy-based selection further increased engraftment. Long-term engraftment and the myeloid-to-lymphoid ratio achieved with vascular niche induction were similar to levels achieved for cord blood-derived MPP and up to 20-fold higher than those achieved with hPSC-derived MPP engraftment. Our findings indicate that endothelial Notch ligands promote PSC-definitive hematopoiesis and production of long-term engrafting CD34(+) cells, suggesting these ligands are critical for HSC emergence.