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A Scalable Suspension Platform for Generating High-Density Cultures of Universal Red Blood Cells from Human Induced Pluripotent Stem Cells.


ABSTRACT: 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.

SUBMITTER: Sivalingam J 

PROVIDER: S-EPMC7897557 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature

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A Scalable Suspension Platform for Generating High-Density Cultures of Universal Red Blood Cells from Human Induced Pluripotent Stem Cells.

Sivalingam Jaichandran J   SuE Yu Y   Lim Zhong Ri ZR   Lam Alan T L ATL   Lee Alison P AP   Lim Hsueh Lee HL   Chen Hong Yu HY   Tan Hong Kee HK   Warrier Tushar T   Hang Jing Wen JW   Nazir Nazmi B NB   Tan Andy H M AHM   Renia Laurent L   Loh Yuin Han YH   Reuveny Shaul S   Malleret Benoit B   Oh Steve K W SKW  

Stem cell reports 20201210 1


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 proce  ...[more]

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