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Brain organoid formation on decellularized porcine brain ECM hydrogels
VERIGRAFT AB, Gothenburg, Sweden / Department of Molecular and Clinical Medicine, Wallenberg Laboratory, University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, Sweden / Department of Biomedical Engineering, Tufts University, Medford, MA, United States of America.
Department of Molecular Biology, Center of Molecular Biology “Severo Ochoa” (UAM-CSIC), Universidad Autonoma de Madrid, Spain.
Department of Biotechnology and Biomedicine, Technical University of Denmark, Kgs. Lyngby, Denmark / FELIXROBOTICS BV, Utrecht, Netherlands.
University of Skövde, School of Bioscience. University of Skövde, Systems Biology Research Environment. (Translationell bioinformatik, Translational Bioinformatics)ORCID iD: 0000-0003-2942-6702
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2021 (English)In: PLOS ONE, E-ISSN 1932-6203, Vol. 16, no 1, article id e0245685Article in journal (Refereed) Published
Abstract [en]

Human brain tissue models such as cerebral organoids are essential tools for developmental and biomedical research. Current methods to generate cerebral organoids often utilize Matrigel as an external scaffold to provide structure and biologically relevant signals. Matrigel however is a nonspecific hydrogel of mouse tumor origin and does not represent the complexity of the brain protein environment. In this study, we investigated the application of a decellularized adult porcine brain extracellular matrix (B-ECM) which could be processed into a hydrogel (B-ECM hydrogel) to be used as a scaffold for human embryonic stem cell (hESC)-derived brain organoids. We decellularized pig brains with a novel detergent- and enzyme-based method and analyzed the biomaterial properties, including protein composition and content, DNA content, mechanical characteristics, surface structure, and antigen presence. Then, we compared the growth of human brain organoid models with the B-ECM hydrogel or Matrigel controls in vitro. We found that the native brain source material was successfully decellularized with little remaining DNA content, while Mass Spectrometry (MS) showed the loss of several brain-specific proteins, while mainly different collagen types remained in the B-ECM. Rheological results revealed stable hydrogel formation, starting from B-ECM hydrogel concentrations of 5 mg/mL. hESCs cultured in B-ECM hydrogels showed gene expression and differentiation outcomes similar to those grown in Matrigel. These results indicate that B-ECM hydrogels can be used as an alternative scaffold for human cerebral organoid formation, and may be further optimized for improved organoid growth by further improving protein retention other than collagen after decellularization.

Place, publisher, year, edition, pages
Public Library of Science , 2021. Vol. 16, no 1, article id e0245685
National Category
Cell and Molecular Biology Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Biochemistry and Molecular Biology
Research subject
Bioinformatics
Identifiers
URN: urn:nbn:se:his:diva-19469DOI: 10.1371/journal.pone.0245685ISI: 000635021400046PubMedID: 33507989Scopus ID: 2-s2.0-85100288123OAI: oai:DiVA.org:his-19469DiVA, id: diva2:1527715
Projects
European Union's Horizon 2020 Research and Innovation Program under the Marie SklodowskaCurie Grant
Funder
EU, Horizon 2020, 722779
Note

CC BY 4.0

© 2021 Simsa et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Available from: 2021-02-11 Created: 2021-02-11 Last updated: 2021-06-14Bibliographically approved

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Ghosheh, Nidal

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