Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA, USA; Department of Surgery, Boston University Medical Center, Boston, MA, USA.
Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA, USA.
Stem Cell Reports. 2022 Dec 13;17(12):2610-2628. doi: 10.1016/j.stemcr.2022.10.007. Epub 2022 Nov 3.
A robust method of producing mature T cells from iPSCs is needed to realize their therapeutic potential. NOTCH1 is known to be required for the production of hematopoietic progenitor cells with T cell potential in vivo. Here we identify a critical window during mesodermal differentiation when Notch activation robustly improves access to definitive hematopoietic progenitors with T/NK cell lineage potential. Low-density progenitors on either OP9-hDLL4 feeder cells or hDLL4-coated plates favored T cell maturation into TCRabCD3CD8 cells that express expected T cell markers, upregulate activation markers, and proliferate in response to T cell stimulus. Single-cell RNAseq shows Notch activation yields a 6-fold increase in multi-potent hematopoietic progenitors that follow a developmental trajectory toward T cells with clear similarity to post-natal human thymocytes. We conclude that early mesodermal Notch activation during hematopoietic differentiation is a missing stimulus with broad implications for producing hematopoietic progenitors with definitive characteristics.
需要一种稳健的方法从 iPS 细胞中产生成熟的 T 细胞，以实现其治疗潜力。已知 NOTCH1 是体内产生具有 T 细胞潜能的造血祖细胞所必需的。在这里，我们确定了中胚层分化过程中的一个关键窗口，在此期间，Notch 激活可有力地提高获得具有 T/NK 细胞谱系潜能的确定性造血祖细胞的能力。在 OP9-hDLL4 饲养细胞或 hDLL4 包被的平板上的低密度祖细胞有利于 T 细胞成熟为 TCRabCD3CD8 细胞，这些细胞表达预期的 T 细胞标记物，上调激活标记物，并对 T 细胞刺激物产生增殖反应。单细胞 RNAseq 显示，Notch 激活可使多能造血祖细胞增加 6 倍，这些祖细胞遵循向具有明显类似于产后人类胸腺细胞特征的 T 细胞发育的轨迹。我们得出结论，造血分化过程中的早期中胚层 Notch 激活是一种缺失的刺激因素，对产生具有确定性特征的造血祖细胞具有广泛的意义。
