The original protocol, which involved the integration of transcription factor combinations into the cell genome, has been refined and now makes use of integration-free factors, which make iPSCs therapy for regenerative medicine applications safer. Since the first report of these cells in 2006, human iPSCs have been derived from many cell types, including blood cells. Induced pluripotent stem cells (iPSCs) are an essentially infinite source of stem cells owing to their self-renewal ability. As human HSCs are a limited resource and ethical concerns hinder the use of embryonic HSCs, the large-scale expansion of RBCs for transfusion purposes remains problematic. There have been numerous advances in the generation of safe erythroid cells and improvements in the efficiency of single hematopoietic cell acquisition by differentiation of CD34 + HSCs using cytokines and small molecules. Hematopoietic stem cells (HSC) obtained from cord blood and bone marrow have been used to produce functional enucleated RBCs. ĭuring the past decade, enormous progress has been made in the ex vivo manufacture of human RBCs. However, these blood substitutes result in insufficient oxygen delivery and an increased likelihood of death, and therefore are not promising alternatives. Despite extensive efforts, it is not always easy to find a suitable blood component, and alternative systems capable of transporting oxygen to the body have been evaluated, including perfluorochemical-based RBC substitutes, hemoglobin-based RBC substitutes, and recombinant hemoglobin. Currently, RBCs are only available from donations by healthy volunteers, but insufficient numbers of donors and the potential for transfusion-transmitted infections remain considerable challenges in meeting the demand for blood. Since the 17th century, the transfusion of red blood cells (RBCs) has been a crucial part of modern medicine, enabling the alleviation of symptoms in patients with severe anemia or trauma. Our results demonstrate the feasibility of producing autologous iPSC-differentiated RBCs for clinical transfusions in patients without alternative options. It has previously only been hypothesized that erythroid differentiation from iPSCs could be used to produce RBCs for transfusion to patients with rare blood types or who have been alloimmunized. This differentiation protocol resulted in moderate erythrocyte yield per iPSC. ResultsĬells from all donors were successfully used to generate iPSC lines, which were differentiated into erythroid precursors without any apparent chromosomal mutations. Morphology and cell counts were determined by microscopy observations and flow cytometry. The stability of iPSC lines was confirmed with chromosomal analysis and RT-PCR. A 31-day serum-free, xeno-free erythroid differentiation protocol was used to generate reticulocytes. Mononuclear cells separated from the peripheral blood of O D-positive and rare blood type donors were cultured to produce and expand erythroid progenitors and reprogrammed into iPSCs. ![]() In this study, we developed a detailed protocol for the generation of iPSC lines derived from peripheral blood of donors with O D-positive blood and rare blood types (D–and Jr(a-)) and subsequent erythroid differentiation. However, limitations like high costs and technological requirements restrict the use of RBCs produced by iPSC differentiation to specific circumstances, such as for patients with rare blood types or alloimmunized patients. The in vitro production of mature human red blood cells (RBCs) from induced pluripotent stem cells (iPSCs) has been the focus of research to meet the high demand for blood transfusions.
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