Abstract
Induced pluripotent stem cells (iPSC) are an attractive progenitor source for the generation of in vitro blood products. However, before iPSC-derived erythroid cells can be considered for therapeutic use their similarity to adult erythroid cells must be confirmed. We have analysed the proteome of erythroid cells differentiated from the iPSC fibroblast derived line (C19) and showed they express hallmark RBC proteins, including all those of the ankyrin and 4.1R complex. We next compared the proteome of erythroid cells differentiated from three iPSC lines (C19, OCE1, OPM2) with that of adult and cord blood progenitors. Of the 1989 proteins quantified <3% differed in level by 2-fold or more between the different iPSC-derived erythroid cells. When compared to adult cells, 11% of proteins differed in level by 2-fold or more, falling to 1.9% if a 5-fold threshold was imposed to accommodate slight inter-cell line erythropoietic developmental variation. Notably, the level of >30 hallmark erythroid proteins was consistent between the iPSC lines and adult cells. In addition, a sub-population (10–15%) of iPSC erythroid cells in each of the iPSC lines completed enucleation. Aberrant expression of some cytoskeleton proteins may contribute to the failure of the majority of the cells to enucleate since we detected some alterations in cytoskeletal protein abundance. In conclusion, the proteome of erythroid cells differentiated from iPSC lines is very similar to that of normal adult erythroid cells, but further work to improve the induction of erythroid cells in existing iPSC lines or to generate novel erythroid cell lines is required before iPSC-derived red cells can be considered suitable for transfusion therapy.
Highlights
The generation of human red blood cells (RBCs) in vitro for transfusion purposes is a major goal of health services globally
The aim of our study was to determine how similar erythroid cells differentiated from induced pluropotent stem cells (iPSC) are to adult erythroid cells generated in vitro, and whether there are any gross differences which could impact on their potential for use as therapeutic source of red cells
Our initial proteome analysis of orthochromatic erythroid cells differentiated from C19 iPSCs revealed no major differences regarding expression of hallmark RBC proteins, all known components of the 4.1R and ankyrin membrane complexes
Summary
The generation of human red blood cells (RBCs) in vitro for transfusion purposes is a major goal of health services globally. The cells are similar to embryonic pluripotent stem cells (ESCs) in their morphology, pluripotency marker expression, self-renewal property and ability to differentiate into the three primary germ layers both in vivo and in vitro [2,3,4,5,6,7]. They do not have the ethical barriers of ESCs [4]. A retrospective study by Peyrard et al (2011) [8] has shown that RBCs generated from only 15 human iPSC lines could cover the needs of patients of Western Caucasians
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