Giorgetti Alessandra, Castaño Julio, Bueno Clara, Díaz de la Guardia Rafael, Delgado Mario, Bigas Anna, Espinosa Lluis, Menendez Pablo
Josep Carreras Leukemia Research Institute and Department of Biomedicine, School of Medicine, University of Barcelona, Barcelona, Spain; Center for Regenerative Medicine in Barcelona (CMRB), Barcelona, Spain.
Josep Carreras Leukemia Research Institute and Department of Biomedicine, School of Medicine, University of Barcelona, Barcelona, Spain.
Exp Hematol. 2017 Jan;45:85-93.e2. doi: 10.1016/j.exphem.2016.09.007. Epub 2016 Sep 28.
Recent studies in zebrafish and mice have revealed that proinflammatory signaling is a positive regulator of definitive hematopoietic development. Whether proinflammatory signaling also regulates human hematopoietic specification remains unknown. Here, we explored the impact of the proinflammatory cytokines tumor necrosis factor-α (TNFα), interferon-γ (IFNγ), and interleukin-1β (IL1β) on in vitro hematopoietic differentiation using human pluripotent stem cells. Gene expression analysis and enzyme-linked immunosorbent assay revealed the absence of a proinflammatory signature during hematopoietic development of human pluripotent stem cells. Functionally, the emergence of hemogenic endothelial progenitors (CD31CD34CD45 or CD34CD43CD73) and hematopoietic cells (CD43CD45) was not affected by treatment with increasing doses of TNFα, IFNγ, and IL1β irrespective of the developmental window or the differentiation protocol used (embryoid body or OP9 co-culture based). Similarly, knockdown of endogenous NF-kB signaling had no impact on hematopoietic differentiation of human pluripotent stem cells. This study serves as a demonstration that TNFα, IFNγ, and IL1β signals do not improve hematopoietic differentiation of human pluripotent stem cells using current protocols and suggests that proinflammatory signaling is insufficient to drive definitive hematopoietic specification of human hematopoietic stem cells in vitro.
最近在斑马鱼和小鼠身上进行的研究表明，促炎信号是确定性造血发育的正向调节因子。促炎信号是否也调节人类造血特化仍不清楚。在此，我们利用人类多能干细胞探索了促炎细胞因子肿瘤坏死因子-α（TNFα）、干扰素-γ（IFNγ）和白细胞介素-1β（IL1β）对体外造血分化的影响。基因表达分析和酶联免疫吸附测定显示，在人类多能干细胞的造血发育过程中不存在促炎特征。在功能上，无论使用何种发育窗口或分化方案（基于胚状体或OP9共培养），用递增剂量的TNFα、IFNγ和IL1β处理均不影响造血内皮祖细胞（CD31CD34CD45或CD34CD43CD73）和造血细胞（CD43CD45）的出现。同样，内源性NF-κB信号的敲低对人类多能干细胞的造血分化没有影响。这项研究表明，使用当前方案，TNFα、IFNγ和IL1β信号不会改善人类多能干细胞的造血分化，并表明促炎信号不足以在体外驱动人类造血干细胞的确定性造血特化。
