Stem Cell Bioprocessing Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, 20 Biopolis Way, Centros 06-01, Singapore 138668, Singapore.
Transcriptomics Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore 138668, Singapore.
Stem Cell Reports. 2021 Jan 12;16(1):182-197. doi: 10.1016/j.stemcr.2020.11.008. Epub 2020 Dec 10.
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 × 10 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.
通用的 O 型阴性人类诱导多能干细胞（hiPSC）分化而来的红细胞（RBC）可应用于输血医学。鉴于每次输血单位需要 2 万亿个 RBC，因此需要开发高效的生物工艺，以便在体外大规模生成 RBC。我们已经开发了一种可扩展的悬浮搅拌培养平台，用于将 hiPSC-微载体聚集体分化为功能性 RBC，并已证明该过程从 6 孔板开始具有可扩展性，最终在 500 mL 搅拌瓶中进行演示。从表现最好的 hiPSC 分化而来的细胞在 50 mL 培养物中产生了 8.5 亿个成红细胞，细胞密度接近 1.7×10^6 个/mL。功能（氧结合、血红蛋白特性、膜完整性和波动）和转录组学评估表明，hiPSC 衍生的 RBC 与成人衍生的 RBC 之间几乎没有差异。我们在这里描述的可扩展搅拌悬浮培养分化过程可应用于未来在受控生物反应器中大规模生产 RBC。
