Sivalingam, J., SuE, Y., Lim, Z. R., Lam, A. T. L., Lee, A. P., Lim, H. L., … Oh, S. K. W. (2021). A Scalable Suspension Platform for Generating High-Density Cultures of Universal Red Blood Cells from Human Induced Pluripotent Stem Cells. Stem Cell Reports, 16(1), 182–197. doi:10.1016/j.stemcr.2020.11.008
Universal red blood cells (RBCs) differentiated from O-negative human induced pluripotent stem cells (hiPSCs) could find applications in transfusion medicine. Given that each transfusion unit of blood requires 2 trillion RBCs, efficient bioprocesses need to be developed for large-scale in vitro generation of RBCs. We have developed a scalable suspension agitation culture platform for differentiating hiPSC-microcarrier aggregates into functional RBCs and have demonstrated scalability of the process starting with 6 well plates and finally demonstrating in 500 mL spinner flasks. Differentiation of the best-performing hiPSCs generated 0.85 billion erythroblasts in 50 mL cultures with cell densities approaching 1.7 × 107 cells/mL. Functional (oxygen binding, hemoglobin characterization, membrane integrity, and fluctuations) and transcriptomics evaluations showed minimal differences between hiPSC-derived and adult-derived RBCs. The scalable agitation suspension culture differentiation process we describe here could find applications in future large-scale production of RBCs in controlled bioreactors.
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This work was supported by a Joint Council Office grant from the Agency for Science, Technology and Research, Singapore (A*STAR) (1331AFG075), Singapore Immunology Network Core funding, A*STAR, and NUHS Start-Up funding (NUHSRO/2018/006/SU/01).