title: In vitro large scale production of human mature red blood cells from hematopoietic stem cells by coculturing with human fetal liver stromal cells.
abstract: In vitro models of human erythropoiesis are useful in studying the mechanisms of erythroid differentiation in normal and pathological conditions. Here we describe an erythroid liquid culture system starting from cord blood derived hematopoietic stem cells (HSCs). HSCs were cultured for more than 50 days in erythroid differentiation conditions and resulted in a more than 10(9)-fold expansion within 50 days under optimal conditions. Homogeneous erythroid cells were characterized by cell morphology, flow cytometry, and hematopoietic colony assays. Furthermore, terminal erythroid maturation was improved by cosculturing with human fetal liver stromal cells. Cocultured erythroid cells underwent multiple maturation events, including decrease in size, increase in glycophorin A expression, and nuclear condensation. This process resulted in extrusion of the pycnotic nuclei in up to 80% of the cells. Importantly, they possessed the capacity to express the adult definitive β -globin chain upon further maturation. We also show that the oxygen equilibrium curves of the cord blood-differentiated red blood cells (RBCs) are comparable to normal RBCs. The large number and purity of erythroid cells and RBCs produced from cord blood make this method useful for fundamental research in erythroid development, and they also provide a basis for future production of available RBCs for transfusion.
authors.affiliation: Stem Cell and Regenerative Medicine Lab, Beijing Institute of Transfusion Medicine, 27 Taiping Road, Beijing 100850, China.
authors.forename: Jiafei
authors.initials: J
authors.lastname: Xi
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Li
authors.forename: Ruoyong
authors.initials: R
authors.lastname: Wang
authors.forename: Yunfang
authors.initials: Y
authors.lastname: Wang
authors.forename: Xue
authors.initials: X
authors.lastname: Nan
authors.forename: Lijuan
authors.initials: L
authors.lastname: He
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.forename: Wen
authors.initials: W
authors.lastname: Yue
authors.forename: Xuetao
authors.initials: X
authors.lastname: Pei
mesh_terms: D018920:Coculture Techniques / Q000379:methods; D004912:Erythrocytes / Q000166:cytology* / Q000378:metabolism; D005260:Female; D005333:Fetus / Q000166:cytology* / Q000378:metabolism; D006021:Glycophorins / Q000096:biosynthesis; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008297:Male; D008954:Models, Biological; D061251:Primary Cell Culture / Q000379:methods*; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism; D055544:beta-Globins / Q000096:biosynthesis
chemical_list: D006021:Glycophorins; D055544:beta-Globins