Extracellular matrix (ECM) plays an important role through structural and biochemical interaction with cells. Tissue-specific ECM, attained through decellularization (decel), has been proposed in many regenerative strategies for tissue and organ replacement. Decel of animal pancreata has been reported, but similar methods applied to human pancreas are less effective due to high lipid content. ECM-derived hydrogel can be made from many tissues, but human pancreas-derived hydrogel has not been reported.

Our objective is to produce an acellular biological scaffold from human pancreas ECM (hP-ECM) for use in tissue culture and transplantation platforms.

Human pancreata (Fig.1a) were decelled with spin and homogenization techniques, using physical and chemical treatments to isolate hP-ECM (1b). hP-ECM was pepsin digested (1c), neutralized and warmed to 37[deg]C to form hP-ECM hydrogel (hP-HG)(1d). hP-ECM and hP-HG were examined for lipid and DNA removal, and retention of ECM protein and glycosaminoglycan (GAG). Cytocompatibility of hP-HG was tested in vitro (1e) and immune response to hP-HG was assessed in vivo (1f).

Lipid content is significantly reduced following homogenize-decel compared to spin-decel; lipid removal significantly enhanced gelation of hP-HG. DNA content is significantly reduced in the hP-ECM (4.2%) and hP-HG (0.44%) compared to the native pancreas (100%) while GAG content is moderately retained in the hP-ECM (20.8%) and hP-HG (4.0%). hP-ECM and hP-HG stain positively for Col1, Col4 and Laminin, but negatively for immunogenic proteins such as HLA Class I and II.

hP-HG is cytocompatible with the beta cell line hINS-1, which grows on hP-HG with equal proficiency as on Col1 or uncoated surfaces. When hPSC-derived pancreatic progenitor cells were embedded in hP-HG the cells retained their Pdx1⁺ fate, were proliferative (Ki67) and had a low apoptosis phenotype (Casp3).

hP-HG transplanted into humanized mice had minimal cellular infiltrate, whereas antigenic tissue was acutely infiltrated by human T cells (CD3) and B cells (CD20).

Conclusion: We developed a novel protocol for the decel of human pancreas and the production of hP-ECM and hP-HG scaffolds suitable for cell culture and transplantation applications.

Fig. 1

CITATION INFORMATION: Tremmel D., Sackett S., Feeney A., Maguire R., Odorico J. Human Pancreatic ECM Scaffolds for Islet Culture and Transplantation Am J Transplant. 2017;17 (suppl 3).

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