Human induced pluripotent stem cell derived erythroblasts can undergo definitive erythropoiesis and co-express gamma and beta globins.
Human induced pluripotent stem cells (hiPSCs), like embryonic stem cells, are under intense investigation for novel approaches to model disease and for regenerative therapies. Here, we describe the derivation and characterization of hiPSCs from a variety of sources and show that, irrespective of origin or method of reprogramming, hiPSCs can be differentiated on OP9 stroma towards a multi-lineage haemo-endothelial progenitor that can contribute to CD144(+) endothelium, CD235a(+) erythrocytes (myeloid lineage) and CD19(+) B lymphocytes (lymphoid lineage). Within the erythroblast lineage, we were able to demonstrate by single cell analysis (flow cytometry), that hiPSC-derived erythroblasts express alpha globin as previously described, and that a sub-population of these erythroblasts also express haemoglobin F (HbF), indicative of fetal definitive erythropoiesis. More notably however, we were able to demonstrate that a small sub-fraction of HbF positive erythroblasts co-expressed HbA in a highly heterogeneous manner, but analogous to cord blood-derived erythroblasts when cultured using similar methods. Moreover, the HbA expressing erythroblast population could be greatly enhanced (44·0 ± 6·04%) when a defined serum-free approach was employed to isolate a CD31(+) CD45(+) erythro-myeloid progenitor. These findings demonstrate that hiPSCs may represent a useful alternative to standard sources of erythrocytes (RBCs) for future applications in transfusion medicine.
Blood Research Laboratory, Radcliffe Department of Medicine, NHS Blood and Transplant and Nuffield Division of Clinical Laboratory Medicine, John Radcliffe Hospital, University of Oxford, Oxford, UK; Stem Cell Research Laboratory, Radcliffe Department of Medicine, NHS Blood and Transplant and Nuffield Division of Clinical Laboratory Medicine, John Radcliffe Hospital, University of Oxford, Oxford, UK.
Cheng-Tao
CT
Yang
Anna
A
French
Pollyanna Agnes
PA
Goh
Alistair
A
Pagnamenta
Sachith
S
Mettananda
Jenny
J
Taylor
Sam
S
Knight
Amit
A
Nathwani
David J
DJ
Roberts
Suzanne M
SM
Watt
Lee
L
Carpenter
D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D056915:DNA Copy Number Variations; D004900:Erythroblasts / Q000166:cytology* / Q000378:metabolism*; D004920:Erythropoiesis / Q000502:physiology*; D015870:Gene Expression; D006801:Humans; D016130:Immunophenotyping; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D007621:Karyotyping; D055544:beta-Globins / Q000235:genetics* / Q000378:metabolism; D055545:gamma-Globins / Q000235:genetics* / Q000378:metabolism
D055544:beta-Globins; D055545:gamma-Globins
erythropoiesis; globin expression; pluripotent stem cell