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Self-organized emergence of hyaline cartilage in hiPSC-derived multi-tissue organoids

Manci Li, Juan E. Abrahante, Amanda Vegoe, Yi Wen Chai, Beth Lindborg, Ferenc Toth, Peter A. Larsen, Timothy D. O’Brien
doi: https://doi.org/10.1101/2021.09.21.461213
Manci Li
1Department of Veterinary and Biomedical Sciences, University of Minnesota, St. Paul, MN 55108
2Minnesota Center for Prion Research and Outreach, College of Veterinary Medicine, St. Paul, University of Minnesota, MN 55108
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  • For correspondence: li000021@umn.edu
Juan E. Abrahante
3University of Minnesota Informatics Institute, University of Minnesota, Minneapolis, MN 55455
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Amanda Vegoe
4Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455
5Department of Veterinary Population Medicine, University of Minnesota, St. Paul, MN 55108
6Sarcio, Inc., Minneapolis, MN 55455
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Yi Wen Chai
4Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455
5Department of Veterinary Population Medicine, University of Minnesota, St. Paul, MN 55108
6Sarcio, Inc., Minneapolis, MN 55455
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Beth Lindborg
4Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455
5Department of Veterinary Population Medicine, University of Minnesota, St. Paul, MN 55108
6Sarcio, Inc., Minneapolis, MN 55455
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Ferenc Toth
7Department of Veterinary Clinical Sciences, University of Minnesota, St. Paul, MN 55108
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Peter A. Larsen
1Department of Veterinary and Biomedical Sciences, University of Minnesota, St. Paul, MN 55108
2Minnesota Center for Prion Research and Outreach, College of Veterinary Medicine, St. Paul, University of Minnesota, MN 55108
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Timothy D. O’Brien
4Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455
5Department of Veterinary Population Medicine, University of Minnesota, St. Paul, MN 55108
6Sarcio, Inc., Minneapolis, MN 55455
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Abstract

Despite holding great therapeutic potential, existing protocols for in vitro chondrogenesis and hyaline cartilage production from human induced pluripotent stem cells (hiPSC) are laborious and complex with unclear long-term consequences. Here, we developed a simple xeno- and feeder-free protocol for human hyaline cartilage production in vitro using hydrogel-cultured multi-tissue organoids (MTOs). We investigate gene regulatory networks during spontaneous hiPSC-MTO differentiation using RNA sequencing and bioinformatic analyses. We find the interplays between BMPs and neural FGF pathways are associated with the phenotype transition of MTOs. We recognize TGF-beta/BMP and Wnt signaling likely contribute to the long-term maintenance of MTO cartilage growth and further adoption of articular cartilage development. By comparing the MTO transcriptome with human lower limb chondrocytes, we observe that the expression of chondrocyte-specific genes in MTO shows a strong correlation with fetal lower limb chondrocytes. Collectively, our findings describe the self-organized emergence of hyaline cartilage in MTO, its associated molecular pathways, and its spontaneous adoption of articular cartilage development trajectory.

Competing Interest Statement

Timothy D. O'Brien, Beth Lindborg, Amanda Vegoe are officers of and hold equity in, Ferenc Toth is a consultant for, and Yi Wen Chai is an employee of, Sarcio Inc., which has an option from the University of Minnesota to commercialize the organoid technology described herein. These interests have been reviewed and managed by the University of Minnesota in accordance with its Conflict of Interest policies. Manci Li and Peter A. Larsen declare no competing interests.

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  • ↵* Co-senior author

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted November 02, 2021.
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Self-organized emergence of hyaline cartilage in hiPSC-derived multi-tissue organoids
Manci Li, Juan E. Abrahante, Amanda Vegoe, Yi Wen Chai, Beth Lindborg, Ferenc Toth, Peter A. Larsen, Timothy D. O’Brien
bioRxiv 2021.09.21.461213; doi: https://doi.org/10.1101/2021.09.21.461213
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Self-organized emergence of hyaline cartilage in hiPSC-derived multi-tissue organoids
Manci Li, Juan E. Abrahante, Amanda Vegoe, Yi Wen Chai, Beth Lindborg, Ferenc Toth, Peter A. Larsen, Timothy D. O’Brien
bioRxiv 2021.09.21.461213; doi: https://doi.org/10.1101/2021.09.21.461213

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