title: Biologic properties and enucleation of red blood cells from human embryonic stem cells.
abstract: Human erythropoiesis is a complex multistep process that involves the differentiation of early erythroid progenitors to mature erythrocytes. Here we show that it is feasible to differentiate and mature human embryonic stem cells (hESCs) into functional oxygen-carrying erythrocytes on a large scale (10(10)-10(11) cells/6-well plate hESCs). We also show for the first time that the oxygen equilibrium curves of the hESC-derived cells are comparable with normal red blood cells and respond to changes in pH and 2,3-diphosphoglyerate. Although these cells mainly expressed fetal and embryonic globins, they also possessed the capacity to express the adult beta-globin chain on further maturation in vitro. Polymerase chain reaction and globin chain specific immunofluorescent analysis showed that the cells increased expression of beta-globin (from 0% to > 16%) after in vitro culture. Importantly, the cells underwent multiple maturation events, including a progressive decrease in size, increase in glycophorin A expression, and chromatin and nuclear condensation. This process resulted in extrusion of the pycnotic nuclei in up to more than 60% of the cells generating red blood cells with a diameter of approximately 6 to 8 mum. The results show that it is feasible to differentiate and mature hESCs into functional oxygen-carrying erythrocytes on a large scale.
authors: Advanced Cell Technology, Worcester, MA, USA.
authors: Shi-Jiang
authors: SJ
authors: Lu
authors: Qiang
authors: Q
authors: Feng
authors: Jennifer S
authors: JS
authors: Park
authors: Loyda
authors: L
authors: Vida
authors: Bao-Shiang
authors: BS
authors: Lee
authors: Michael
authors: M
authors: Strausbauch
authors: Peter J
authors: PJ
authors: Wettstein
authors: George R
authors: GR
authors: Honig
authors: Robert
authors: R
authors: Lanza
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000502:physiology*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D041905:Erythroid Cells / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D051379:Mice; D012204:Rh-Hr Blood-Group System / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods
chemical_list: D012204:Rh-Hr Blood-Group System; C026410:Rho(D) antigen