Student Catholic University of Korea Catholic University of Korea, Republic of Korea
Abstract: Blood transfusions play a crucial role in modern medicine, but donated blood comes with challenges such as donor shortage and risk of disease transmission. Therefore, the technology of producing red blood cells in vitro with the same quality and function as natural red blood cells (RBC) is an attractive concept and seems indispensable. Human induced pluripotent stem cells(hiPSCs) are emerging as an alternative to overcome donation-dependent transfusions because they have less limitations in cell supply and can differentiate into mature erythrocytes in vitro in a laboratory setting. However, the production and expansion of pluripotent hematopoietic stem/progenitor cells (HPSCs) for clinical application remains a challenge. In particular, the use of feeder cells, such as OP9 cells, to achieve high-efficiency RBC differentiation carries a risk of contamination by heterologous pathogens during culture or in medium, which consequently limits its clinical use. In this study, efficiency was directly compared and analyzed using two experimental methods in the process of producing RBC from hiPSC without stromal cells; dissociation and three-dimensional(3D) culture. Our results confirmed that 3D cultures had a relatively higher efficiency in differentiating RBCs compared to dissociation, and therefore, we established a relatively simple and highly efficient protocol for differentiating RBCs from hiPSCs without using stromal cells. Our findings suggest the possibility of artificial blood production for future clinical transfusions.
Funding Source: This work was supported by the National Research Foundation of Korea (NRF) and ministries including Science and ICT, Trade, Industry and Energy, Health & Welfare, and Food and Drug Safety (RS-2023-KH142779, RS-2024-00512348).
