Extracellular matrix scaffold and hydrogel derived from decellularized and delipidized human pancreas

TLDR

Extracellular matrix (ECM) regulates cell behavior, and tissue‑specific ECM obtained by decellularization has been explored for organ replacement; however, human pancreas decellularization is challenged by high lipid content, and no protocols for deriving a pancreas‑derived hydrogel have been reported. The study aims to develop and characterize a novel decellularization and delipidization protocol for human pancreas to produce acellular ECM and hydrogel suitable for cell culture and transplantation. The authors applied novel decellularization techniques to human pancreas, yielding an acellular 3D scaffold (hP‑ECM) and hydrogel (hP‑HG) suitable for culture, transplantation, and proteomic studies. The protocol, incorporating a homogenization step, markedly enhanced lipid removal and enabled the ECM to gel at 37 °C both in vitro and in vivo, proved cytocompatible with diverse cell types and islet‑like tissues, and revealed 120 proteins in the human pancreatic matrisome.

Abstract

Extracellular matrix (ECM) plays an important developmental role by regulating cell behaviour through structural and biochemical stimulation. Tissue-specific ECM, attained through decellularization, has been proposed in several strategies for tissue and organ replacement. Decellularization of animal pancreata has been reported, but the same methods applied to human pancreas are less effective due to higher lipid content. Moreover, ECM-derived hydrogels can be obtained from many decellularized tissues, but methods have not been reported to obtain human pancreas-derived hydrogel. Using novel decellularization methods with human pancreas we produced an acellular, 3D biological scaffold (hP-ECM) and hydrogel (hP-HG) amenable to tissue culture, transplantation and proteomic applications. The inclusion of a homogenization step in the decellularization protocol significantly improved lipid removal and gelation capability of the resulting ECM, which was capable of gelation at 37 °C in vitro and in vivo, and is cytocompatible with a variety of cell types and islet-like tissues in vitro. Overall, this study demonstrates the characterisation of a novel protocol for the decellularization and delipidization of human pancreatic tissue for the production of acellular ECM and ECM hydrogel suitable for cell culture and transplantation applications. We also report a list of 120 proteins present within the human pancreatic matrisome.