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  • In-Depth Analysis of the Pancreatic Extracellular Matrix during Development for Next-Generation Tissue Engineering.

In-Depth Analysis of the Pancreatic Extracellular Matrix during Development for Next-Generation Tissue Engineering.

International journal of molecular sciences (2023-06-28)
Laura Glorieux, Laura Vandooren, Sylvie Derclaye, Sébastien Pyr Dit Ruys, Paloma Oncina-Gil, Anna Salowka, Gaëtan Herinckx, Elias Aajja, Pascale Lemoine, Catherine Spourquet, Hélène Lefort, Patrick Henriet, Donatienne Tyteca, Francesca M Spagnoli, David Alsteens, Didier Vertommen, Christophe E Pierreux
ABSTRACT

The pancreas is a complex organ consisting of differentiated cells and extracellular matrix (ECM) organized adequately to enable its endocrine and exocrine functions. Although much is known about the intrinsic factors that control pancreas development, very few studies have focused on the microenvironment surrounding pancreatic cells. This environment is composed of various cells and ECM components, which play a critical role in maintaining tissue organization and homeostasis. In this study, we applied mass spectrometry to identify and quantify the ECM composition of the developing pancreas at the embryonic (E) day 14.5 and postnatal (P) day 1 stages. Our proteomic analysis identified 160 ECM proteins that displayed a dynamic expression profile with a shift in collagens and proteoglycans. Furthermore, we used atomic force microscopy to measure the biomechanical properties and found that the pancreatic ECM was soft (≤400 Pa) with no significant change during pancreas maturation. Lastly, we optimized a decellularization protocol for P1 pancreatic tissues, incorporating a preliminary crosslinking step, which effectively preserved the 3D organization of the ECM. The resulting ECM scaffold proved suitable for recellularization studies. Our findings provide insights into the composition and biomechanics of the pancreatic embryonic and perinatal ECM, offering a foundation for future studies investigating the dynamic interactions between the ECM and pancreatic cells.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Laminin antibody produced in rabbit, 0.5 mg/mL, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Papain from papaya latex, buffered aqueous suspension, 2× Crystallized, ≥16 units/mg protein
Sigma-Aldrich
L-Cysteine hydrochloride, anhydrous, ≥99.0% (RT)