Large-Scale Production of Wholly Cellular Bioinks via the Optimization of Human Induced Pluripotent Stem Cell Aggregate Culture in Automated Bioreactors.

Autor: Ho DLL; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA., Lee S; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA., Du J; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA., Weiss JD; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA., Tam T; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA., Sinha S; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA., Klinger D; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA., Devine S; Sartorius Stedim North America Inc, 565 Johnson Avenue, Bohemia, NY, 11716, USA., Hamfeldt A; Sartorius Stedim North America Inc, 565 Johnson Avenue, Bohemia, NY, 11716, USA., Leng HT; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA., Herrmann JE; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA.; School of Medicine, Stanford University, Stanford, CA, 94305, USA., He M; Materials Science and Engineering, Stanford University, Stanford, CA, 94305, USA., Fradkin LG; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA., Tan TK; Institute of Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, CA, 94305, USA.; Department of Genetics, Stanford University School of Medicine, Stanford, CA, 94305, USA.; Department of Pathology, Stanford University School of Medicine, Stanford, CA, 94305, USA., Standish D; Sartorius Stedim North America Inc, 565 Johnson Avenue, Bohemia, NY, 11716, USA., Tomasello P; Sartorius Stedim North America Inc, 565 Johnson Avenue, Bohemia, NY, 11716, USA., Traul D; Sartorius Stedim North America Inc, 565 Johnson Avenue, Bohemia, NY, 11716, USA., Dianat N; Sartorius Stedim France S.A.S, Zone Industrielle les Paluds, Avenue de Jouques CS 71058, Aubagne Cedex, 13781, France., Ladi R; Sartorius Stedim North America Inc, 565 Johnson Avenue, Bohemia, NY, 11716, USA., Vicard Q; Sartorius Stedim France S.A.S, Zone Industrielle les Paluds, Avenue de Jouques CS 71058, Aubagne Cedex, 13781, France., Katikireddy K; Sartorius Stedim North America Inc, 565 Johnson Avenue, Bohemia, NY, 11716, USA., Skylar-Scott MA; Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA.; Basic Science and Engineering Initiative, Children's Heart Center, Stanford University, Stanford, CA, 94305, USA.; Chan Zuckerberg Biohub, San Francisco, CA, 94158, USA.
Jazyk: angličtina
Zdroj: Advanced healthcare materials [Adv Healthc Mater] 2022 Dec; Vol. 11 (24), pp. e2201138. Date of Electronic Publication: 2022 Nov 22.
DOI: 10.1002/adhm.202201138
Abstrakt: 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.
(© 2022 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.)
Databáze: MEDLINE
Externí odkaz: