Induced pluripotent stem cells (iPSCs) are an invaluable resource for the study of human disease. However, there are no standardized methods for differentiation into hematopoietic cells, and there is a lack of robust, direct comparisons of different methodologies. In the current study we improved a feeder-free, serum-free method for generation of hematopoietic cells from iPSCs, and directly compared this with three other commonly used strategies with respect to efficiency, repeatability, hands-on time, and cost. We also investigated their capability and sensitivity to model genetic hematopoietic disorders in cells derived from Down syndrome and β-thalassemia patients. Of these methods, a multistep monolayer-based method incorporating aryl hydrocarbon receptor hyperactivation (“2D-multistep”) was the most efficient, generating significantly higher numbers of CD34⁺ progenitor cells and functional hematopoietic progenitors, while being the most time- and cost-effective and most accurately recapitulating phenotypes of Down syndrome and β-thalassemia.

诱导多能干细胞（iPSC）是研究人类疾病的宝贵资源。然而，尚无用于分化为造血细胞的标准化方法，并且缺乏对不同方法的可靠，直接的比较。在当前研究中，我们改进了从iPSC生成造血细胞的无饲养层，无血清方法，并将其与其他三种常用策略在效率，可重复性，动手时间和成本方面进行了直接比较。我们还研究了它们在唐氏综合症和β地中海贫血患者的细胞中建模遗传性造血疾病的能力和敏感性。在这些方法中，最有效的方法是结合芳基烃受体超活化的基于多步单层的方法（“ 2D-multistep”），⁺祖细胞和功能性造血祖细胞，同时是唐氏综合症和β地中海贫血的最省时，最具成本效益和最准确的概括表型。