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Biomaterials. 2018 Sep;177:27-39. doi: 10.1016/j.biomaterials.2018.05.031. Epub 2018 May 25.

3D heterogeneous islet organoid generation from human embryonic stem cells using a novel engineered hydrogel platform.

Author information

1
Department of Bioengineering, University of Pittsburgh, PA, United States.
2
Biomedical Engineering, Arizona State University, Tempe, AZ, United States.
3
Department of Chemical and Petroleum Engineering, University of Pittsburgh, PA, United States.
4
Department of Physics, Center for Biological Physics, and Biodesign Institute, Arizona State University, Tempe, AZ, United States.
5
Department of Bioengineering, University of Pittsburgh, PA, United States; Department of Chemical and Petroleum Engineering, University of Pittsburgh, PA, United States; Department of Mechanical Engineering and Material Science, University of Pittsburgh, PA, United States; Center for Complex Engineered Multifunctional Materials, University of Pittsburgh, PA, United States; McGowan Institute for Regenerative Medicine, University of Pittsburgh, PA, United States.
6
Chemical Engineering, Arizona State University, Tempe, AZ, United States.
7
Department of Bioengineering, University of Pittsburgh, PA, United States; Department of Chemical and Petroleum Engineering, University of Pittsburgh, PA, United States; McGowan Institute for Regenerative Medicine, University of Pittsburgh, PA, United States. Electronic address: ipb1@pitt.edu.

Abstract

Organoids, which exhibit spontaneous organ specific organization, function, and multi-cellular complexity, are in essence the in vitro reproduction of specific in vivo organ systems. Recent work has demonstrated human pluripotent stem cells (hPSCs) as a viable regenerative cell source for tissue-specific organoid engineering. This is especially relevant for engineering islet organoids, due to the recent advances in generating functional beta-like cells from human pluripotent stem cells. In this study, we report specific engineering of regenerative islet organoids of precise size and cellular heterogeneity, using a novel hydrogel system, Amikagel. Amikagel facilitated controlled and spontaneous aggregation of human embryonic stem cell derived pancreatic progenitor cells (hESC-PP) into robust homogeneous spheroids. This platform further allowed fine control over the integration of multiple cell populations to produce heterogeneous spheroids, which is a necessity for complex organoid engineering. Amikagel induced hESC-PP spheroid formation enhanced pancreatic islet-specific Pdx-1 and NKX6.1 gene and protein expression, while also increasing the percentage of committed population. hESC-PP spheroids were further induced towards mature beta-like cells which demonstrated increased Beta-cell specific INS1 gene and C-peptide protein expression along with functional insulin production in response to in vitro glucose challenge. Further integration of hESC-PP with biologically relevant supporting endothelial cells resulted in multicellular organoids which demonstrated spontaneous maturation towards islet-specific INS1 gene and C-peptide protein expression along with a significantly developed extracellular matrix support system. These findings establish Amikagel -facilitated platform ideal for islet organoid engineering.

KEYWORDS:

Aggregation; Human embryonic stem cells; Hydrogel; Islet; Organoid; Three dimensional

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