Large‐Scale Production of Wholly Cellular Bioinks via the Optimization of Human Induced Pluripotent Stem Cell Aggregate Culture in Automated Bioreactors. Issue 24 (22nd November 2022)

• Record Type: Journal Article
• Authors: Ho, Debbie L. L.; Lee, Stacey; Du, Jianyi; Weiss, Jonathan D.; Tam, Tony; Sinha, Soham; Klinger, Danielle; Devine, Sean; Hamfeldt, Art; Leng, Hope T.; Herrmann, Jessica E.; He, Mengdi; Fradkin, Lee G.; Tan, Tze Kai; Standish, David; Tomasello, Peter; Traul, Donald; Dianat, Noushin; Ladi, Rukmini; Vicard, Quentin; Katikireddy, Kishore; Skylar‐Scott, Mark A.
• Other Names: Rnjak‐Kovacina Jelena guestEditor.; Choi Yu Suk guestEditor.; Lim Khoon S. guestEditor.
• Is Part Of: Advanced healthcare materials. Volume 11:Issue 24(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-11-22
• Subjects: 3D bioprinting -- cell manufacturing -- organoids -- pluripotent stem cells -- suspension culture; Biomedical materials -- Periodicals; 610.28
• DOI: 10.1002/adhm.202201138
• Languages: English
• ISSNs: 2192-2640

Abstract

Abstract: Combining the sustainable culture of billions of human cells and the bioprinting of wholly cellular bioinks offers a pathway toward organ‐scale tissue engineering. Traditional 2D culture methods are not inherently scalable due to cost, space, and handling constraints. Here, the suspension culture of human induced pluripotent stem cell‐derived aggregates (hAs) is optimized using an automated 250 mL stirred tank bioreactor system. Cell yield, aggregate morphology, and pluripotency marker expression are maintained over three serial passages in two distinct cell lines. Furthermore, it is demonstrated that the same optimized parameters can be scaled to an automated 1 L stirred tank bioreactor system. This 4‐day culture results in a 16.6‐ to 20.4‐fold expansion of cells, generating approximately 4 billion cells per vessel, while maintaining >94% expression of pluripotency markers. The pluripotent aggregates can be subsequently differentiated into derivatives of the three germ layers, including cardiac aggregates, and vascular, cortical and intestinal organoids. Finally, the aggregates are compacted into a wholly cellular bioink for rheological characterization and 3D bioprinting. The printed hAs are subsequently differentiated into neuronal and vascular tissue. This work demonstrates an optimized suspension culture‐to‐3D bioprinting pipeline that enables a sustainable approach to billion cell‐scale organ engineering. Abstract : Practicing the art of manufacturing densely cellular solid organs on demand will require the production of billions of stem cells. Here, an optimized pipeline is presented for growing up to 4 billion human induced pluripotent stem cells in automated bioreactor systems. These stem cell aggregates are compacted into viscoelastic and wholly cellular bioinks and are subsequently bioprinted and differentiated.