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RESEARCH ARTICLE

Toward robust and reproducible pluripotent stem cell expansion in bioprinted GelMA constructs

Elizabeth R Komosa1,2 ,  Wei-Han Lin1,2 ,  Brenda M Ogle1,2,3,4,5,6 *
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1 Department of Biomedical Engineering, College of Science and Engineering, University of Minnesota, Minneapolis, Minnesota, United States of America
2 Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, United States of America
3 Lillehei Heart Institute, University of Minnesota, Minneapolis, Minnesota, United States of America
4 Department of Pediatrics, Medical School, University of Minnesota, Minneapolis, Minnesota, United States of America
5 Institute for Engineering in Medicine, University of Minnesota, Minneapolis, Minnesota, United States of America
6 Masonic Cancer Center, University of Minnesota, Minneapolis, MN, United States of America
IJB, 4633 https://doi.org/10.36922/ijb.4633
Submitted: 22 August 2024 | Accepted: 9 December 2024 | Published: 10 December 2024
(This article belongs to the Special Issue Bioprinting for Tissue Engineering and Modeling)
© 2024 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Combining the technologies of 3D bioprinting and human induced pluripotent stem cells (hiPSCs) has allowed for the creation of tissues with organ-level function in the lab, a promising technique for disease modeling and regenerative medicine. Expanding the stem cells in bioprinted tissues prior to differentiation allows for high cell density, which is important for the formation of cell-cell junctions necessary for macroscale function upon differentiation. Yet, stem cell expansion, critical to successful in situ differentiation, depends heavily on the composition of the bioprinted scaffold. Here, we demonstrate how a common bioink component, gelatin methacryloyl (GelMA), varies depending on the vendor and degree of functionalization. We found that the vendor/GelMA production technique played a greater role in dictating the mechanical properties of the bioprinted constructs than the degree of functionalization, emphasizing the importance of reporting detailed characterization of GelMA scaffolds. Furthermore, the ability of singularized hiPSCs to survive and expand in GelMA scaffolds greatly varied across batches from different vendors and degrees of functionalization, where expansion correlated with the mechanical properties of the scaffold. Yet, we found that using a commercial cloning supplement could restore the ability of single hiPSCs to survive and expand across GelMA types, thus compensating for the varied mechanical properties of the scaffolds. T hese fi ndings provide a pr actical guide fo r the ex pansion of hiPSCs in GelMA constructs with various mechanical properties as required for successful in situ differentiation.

 

Graphical abstract
Keywords
Bioprinting
Scaffold characterization
GelMA
Human induced pluripotent stem cells
In situ expansion
Funding
This work was supported by the EK, Grant 905669. National Institutes of Health, National Heart, Lung, and Blood Institute to BMO, R01 HL160779.
Conflict of interest
The authors declare they have no competing interests.
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International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing
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