造血分化
Hematopoietic differentiation
作者信息
Choi Kyung-Dal, Vodyanik Maxim, Slukvin Igor I
机构信息
Department of Pathology and Laboratory Medicine, University of Wisconsin, Madison, WI 53715 US
A protocol for hematopoietic differentiation of human pluripotent stem cells (hPSCs) and generation of mature myeloid cells from hPSCs through expansion and differentiation of hPSC-derived linCD34CD43CD45 multipotent progenitors. The protocol comprises three major steps: (i) induction of hematopoietic differentiation by coculture of hPSCs with OP9 bone marrow stromal cells; (ii) short-term expansion of multipotent myeloid progenitors with a high dose of granulocyte-macrophage colony-stimulating factor; and (iii) directed differentiation of myeloid progenitors into neutrophils, eosinophils, dendritic cells, Langerhans cells, macrophages and osteoclasts. The generation of multipotent hematopoietic progenitors from hPSCs requires 9 d of culture and an additional 2 d to expand myeloid progenitors. Differentiation of myeloid progenitors into mature myeloid cells requires an additional 5–19 d of culture with cytokines, depending on the cell type.
一种用于人类多能干细胞（hPSC）造血分化以及通过hPSC来源的lin⁻CD34⁺CD43⁻CD45⁺多能祖细胞的扩增和分化从hPSC生成成熟髓细胞的方案。该方案包括三个主要步骤：（i）通过hPSC与OP9骨髓基质细胞共培养诱导造血分化；（ii）用高剂量粒细胞-巨噬细胞集落刺激因子短期扩增多能髓祖细胞；（iii）将髓祖细胞定向分化为中性粒细胞、嗜酸性粒细胞、树突状细胞、朗格汉斯细胞、巨噬细胞和破骨细胞。从hPSC生成多能造血祖细胞需要9天的培养时间，另外还需要2天来扩增髓祖细胞。根据细胞类型，将髓祖细胞分化为成熟髓细胞还需要在细胞因子存在的情况下额外培养5 - 19天。
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