Establishment of Immortalized Human Erythroid Progenitor Cell Lines Able to Produce Enucleated Red Blood Cells

Transfusion of red blood cells (RBCs) is a standard and indispensable therapy in current clinical practice. In vitro production of RBCs offers a potential means to overcome a shortage of transfusable RBCs in some clinical situations and also to provide a source of cells free from possible infection or contamination by microorganisms. Thus, in vitro production of RBCs may become a standard procedure in the future. We previously reported the successful establishment of immortalized mouse erythroid progenitor cell lines that were able to produce mature RBCs very efficiently. Here, we have developed a reliable protocol for establishing immortalized human erythroid progenitor cell lines that are able to produce enucleated RBCs. These immortalized cell lines produce functional hemoglobin and express erythroid-specific markers, and these markers are upregulated following induction of differentiation in vitro. Most importantly, these immortalized cell lines all produce enucleated RBCs after induction of differentiation in vitro, although the efficiency of producing enucleated RBCs remains to be improved further. To the best of our knowledge, this is the first demonstration of the feasibility of using immortalized human erythroid progenitor cell lines as an ex vivo source for production of enucleated RBCs.红细胞（red blood cells, RBCs）输注是当前临床实践中标准且不可或缺的治疗手段。体外（in vitro）制备红细胞可为缓解部分临床场景下可输注红细胞短缺问题提供潜在方案，同时可获得不受微生物感染或污染风险影响的细胞来源，因此有望在未来成为标准化临床操作流程。我们此前已成功建立可高效产生成熟红细胞的永生化小鼠红系祖细胞（erythroid progenitor cell）系。本研究开发了一套可靠的实验方案，用于构建能够产生去核红细胞（enucleated RBCs）的永生化人红系祖细胞系。此类永生化细胞系可表达功能性血红蛋白，并呈现红系特异性标志物；在体外诱导分化后，这些标志物的表达水平会上调。尤为关键的是，尽管去核红细胞的生产效率仍有待进一步提升，但所有此类永生化细胞系在体外诱导分化后均可产生去核红细胞。据我们所知，本研究首次证实了以永生化人红系祖细胞系作为离体（ex vivo）来源生产去核红细胞的可行性。