Laboratory of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan.
Laboratory of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan ; Clinical Application Department, Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan.
Stem Cell Reports. 2013 Dec 5;1(6):499-508. doi: 10.1016/j.stemcr.2013.10.010. eCollection 2013.
The lack of knowledge about the mechanism of erythrocyte biogenesis through self-replication makes the in vitro generation of large quantities of cells difficult. We show that transduction of c-MYC and BCL-XL into multipotent hematopoietic progenitor cells derived from pluripotent stem cells and gene overexpression enable sustained exponential self-replication of glycophorin A(+) erythroblasts, which we term immortalized erythrocyte progenitor cells (imERYPCs). In an inducible expression system, turning off the overexpression of c-MYC and BCL-XL enabled imERYPCs to mature with chromatin condensation and reduced cell size, hemoglobin synthesis, downregulation of GCN5, upregulation of GATA1, and endogenous BCL-XL and RAF1, all of which appeared to recapitulate normal erythropoiesis. imERYPCs mostly displayed fetal-type hemoglobin and normal oxygen dissociation in vitro and circulation in immunodeficient mice following transfusion. Using critical factors to induce imERYPCs provides a model of erythrocyte biogenesis that could potentially contribute to a stable supply of erythrocytes for donor-independent transfusion.
由于缺乏对红细胞自我复制机制的了解，使得大量细胞的体外生成变得困难。我们发现，将 c-MYC 和 BCL-XL 转导到多能造血祖细胞中，并通过基因过表达，可以使糖蛋白 A(+)红细胞持续进行指数级自我复制，我们将其称为永生化红细胞祖细胞（imERYPC）。在一个可诱导表达系统中，关闭 c-MYC 和 BCL-XL 的过表达，可使 imERYPC 成熟，表现为染色质浓缩和细胞体积减小、血红蛋白合成减少、GCN5 下调、GATA1 上调以及内源性 BCL-XL 和 RAF1 上调，所有这些都似乎再现了正常的红细胞生成。imERYPC 主要显示胎儿型血红蛋白，并且在体外和免疫缺陷小鼠的循环中具有正常的氧离解能力。使用关键因子诱导 imERYPC，可以为红细胞生成提供模型，从而可能有助于稳定供应用于非供体输血的红细胞。
