TY - JOUR

T1 - Engineering an endothelialized, endocrine Neo-Pancreas

T2 - Evaluation of islet functionality in an ex vivo model

AU - Everwien, Hannah

AU - Keshi, Eriselda

AU - Hillebrandt, Karl H.

AU - Ludwig, Barbara

AU - Weinhart, Marie

AU - Tang, Peter

AU - Beierle, Anika S.

AU - Napierala, Hendrik

AU - Gassner, Joseph MGV

AU - Seiffert, Nicolai

AU - Moosburner, Simon

AU - Geisel, Dominik

AU - Reutzel-Selke, Anja

AU - Strücker, Benjamin

AU - Pratschke, Johann

AU - Haep, Nils

AU - Sauer, Igor M.

N1 - Funding Information: Hannah Everwien was a participant in the BIH-Medical Doctoral Research Stipends program funded by the Charit? ? Universit?tsmedizin Berlin and the Berlin Institute of Health. Dr. K. Hillebrandt is participant in the BIH-Charit? Junior Clinician Scientist Program funded by the Charit?-Universit?tsmedizin Berlin and the Berlin Institute of Health. Priv.-Doz. Dr. B. Struecker was participant in the BIH?Charit? Clinician Scientist Program funded by the Charit? ? Universit?tsmedizin Berlin and the Berlin Institute of Health. Furthermore, we gratefully thank Prof. Dr. S. Bachmann and Mrs. P. Schrade (both Institute of Vegetative Anatomy, Charit? ? Universit?tsmedizin Berlin) for their help. We also thank D. Wyrwal (Department of Surgery, Charit? ? Universit?tsmedizin Berlin) for his help and support during the project. The authors acknowledge the support of the Cluster of Excellence ?Matters of Activity. Image Space Material? funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany's Excellence Strategy ? EXC 2025 ? 390648296. Publisher Copyright: © 2020

PY - 2020/11

Y1 - 2020/11

N2 - Islet-based recellularization of decellularized, repurposed rat livers may form a transplantable Neo-Pancreas. The aim of this study is the establishment of the necessary protocols, the evaluation of the organ structure and the analysis of the islet functionality ex vivo. After perfusion-based decellularization of rat livers, matrices were repopulated with endothelial cells and mesenchymal stromal cells, incubated for 8 days in a perfusion chamber, and finally repopulated on day 9 with intact rodent islets. Integrity and quality of re-endothelialization was assessed by histology and FITC-dextran perfusion assay. Functionality of the islets of Langerhans was determined on day 10 and day 12 via glucose stimulated insulin secretion. Blood gas analysis variables confirmed the stability of the perfusion cultivation. Histological staining showed that cells formed a monolayer inside the intact vascular structure. These findings were confirmed by electron microscopy. Islets infused via the bile duct could histologically be found in the parenchymal space. Adequate insulin secretion after glucose stimulation after 1-day and 3-day cultivation verified islet viability and functionality after the repopulation process. We provide the first proof-of-concept for the functionality of islets of Langerhans engrafted in a decellularized rat liver. Furthermore, a re-endothelialization step was implemented to provide implantability. This technique can serve as a bioengineered platform to generate implantable and functional endocrine Neo-Pancreases.

AB - Islet-based recellularization of decellularized, repurposed rat livers may form a transplantable Neo-Pancreas. The aim of this study is the establishment of the necessary protocols, the evaluation of the organ structure and the analysis of the islet functionality ex vivo. After perfusion-based decellularization of rat livers, matrices were repopulated with endothelial cells and mesenchymal stromal cells, incubated for 8 days in a perfusion chamber, and finally repopulated on day 9 with intact rodent islets. Integrity and quality of re-endothelialization was assessed by histology and FITC-dextran perfusion assay. Functionality of the islets of Langerhans was determined on day 10 and day 12 via glucose stimulated insulin secretion. Blood gas analysis variables confirmed the stability of the perfusion cultivation. Histological staining showed that cells formed a monolayer inside the intact vascular structure. These findings were confirmed by electron microscopy. Islets infused via the bile duct could histologically be found in the parenchymal space. Adequate insulin secretion after glucose stimulation after 1-day and 3-day cultivation verified islet viability and functionality after the repopulation process. We provide the first proof-of-concept for the functionality of islets of Langerhans engrafted in a decellularized rat liver. Furthermore, a re-endothelialization step was implemented to provide implantability. This technique can serve as a bioengineered platform to generate implantable and functional endocrine Neo-Pancreases.

KW - Diabetes mellitus

KW - Islets of Langerhans

KW - Neo-Pancreas

KW - Re-endothelialization

KW - Tissue engineering

UR - http://www.scopus.com/inward/record.url?scp=85091234388&partnerID=8YFLogxK

U2 - 10.1016/j.actbio.2020.09.022

DO - 10.1016/j.actbio.2020.09.022

M3 - Article

C2 - 32949822

AN - SCOPUS:85091234388

VL - 117

SP - 213

EP - 225

JO - Acta Biomaterialia

JF - Acta Biomaterialia

SN - 1742-7061

ER -