Immortalization of Erythroblasts by c-MYC and BCL-XL Enables Large-Scale Erythrocyte Production from Human Pluripotent Stem Cells

Abstract

SummaryThe 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.

Erythroid differentiation of human induced pluripotent stem cells is independent of donor cell type of origin

“…Strategies to improve expansion rates might focus on increased induction of early hematopoietic and erythroid progenitors from human iPSC and the maximization Epigenetic memory in iPSC and erythropoiesis haematologica | 2015; 100 (1)of their subsequent expansion (e.g. by controlled expression of erythroid-specific genes, introduction of proliferationassociated transcription factors, 47,48 modification of growth factor combinations or mimicry of endogenous niches). Once all these difficulties have been successfully overcome, ex vivo manufactured RBC from human iPSC might be used for personalized transfusion medicine, preferentially applied to allo-immunized patients or those with rare blood groups.…”

Effect of Blood Transfusion on Supportive Therapy of Elderly Patients at Kumamoto, Japan as Compared with Khon Kaen, Thailand

“…In fact, number of blood donation by age group changed dramatically within 10 years between 2003 and 2012 [14]. The number in the 3rd decade decreased from 1,480,855 (26.3%) to 1,000,086 (19.0%) and the number in the 4th decade decreased from 1,470,298 (26.2%) to 1,243,040 (23.6%).…”

Ex vivo Generation of Genetically Modified Macrophages from Human Induced Pluripotent Stem Cells

“…The efficient generation of functional mature myeloid cells has been described by several groups utilizing different strategies, which include embryoid body formation, coculture systems as well as small molecule-or cytokine-driven approaches [3][4][5][6]. In this context, the scalable production of erythrocytes or megakaryocytes/platelets is of particular interest for transfusion medicine [7,8]. Another highly attractive target population for cell-therapeutic strategies is represented by monocyte/macrophages, including tissue-resident macrophages (TRMs).…”