Membrane Properties of Human Induced Pluripotent Stem Cell-Derived Cultured Red Blood Cells

Abstract

Cultured red blood cells from human induced pluripotent stem cells (cRBC_iPSCs) are a promising source for future concepts in transfusion medicine. Before cRBC_iPSCs will have entrance into clinical or laboratory use, their functional properties and safety have to be carefully validated. Due to the limitations of established culture systems, such studies are still missing. Improved erythropoiesis in a recently established culture system, closer simulating the physiological niche, enabled us to conduct functional characterization of enucleated cRBC_iPSCs with a focus on membrane properties. Morphology and maturation stage of cRBC_iPSCs were closer to native reticulocytes (nRETs) than to native red blood cells (nRBCs). Whereas osmotic resistance of cRBC_iPSCs was similar to nRETs, their deformability was slightly impaired. Since no obvious alterations in membrane morphology, lipid composition, and major membrane associated protein patterns were observed, reduced deformability might be caused by a more primitive nature of cRBC_iPSCs comparable to human embryonic- or fetal liver erythropoiesis. Blood group phenotyping of cRBC_iPSCs further confirmed the potency of cRBC_iPSCs as a prospective device in pre-transfusional routine diagnostics. Therefore, RBC membrane analyses obtained in this study underscore the overall prospects of cRBC_iPSCs for their future application in the field of transfusion medicine.

Generation of red blood cells from stem cells: Achievements, opportunities and perspectives for malaria research

“…The widely accepted endpoint of most successful in vitro erythroid culture systems is the youngest class of circulating red blood cells, otherwise known as reticulocytes ( Giarratana et al., 2011 ; Griffiths et al., 2012 ; Shah et al., 2014 ; Heshusius et al., 2019 ; Pellegrin et al., 2021 ; Bernecker et al., 2022 ). The immediate precursor to the mature biconcave erythrocyte, the term reticulocyte describes enucleated cells (of evolving maturity) that are generated following extrusion of the erythroblast condensed nucleus (pyrenocyte) and in vivo exit the bone marrow to enter the bloodstream and circulate the body, remodelling their membrane to achieve biconcavity as they do so.…”