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In Vitro Regeneration of Decellularized Pig Esophagus Using Human Amniotic Stem Cells

Journal article
Authors Nikhil Nayakawde
Ketaki Methe
Debashish Banerjee
Malin Berg
Goditha U Premaratne
Michael Olausson
Published in Bioresearch Open Access
Volume 9
Issue 1
Pages 22-36
ISSN 2164-7860
Publication year 2020
Published at Institute of Clinical Sciences, Department of Surgery
Institute of Medicine
Institute of Clinical Sciences, Department of Otorhinolaryngology
Pages 22-36
Language en
Links dx.doi.org/10.1089/biores.2019.0054
Keywords cell adhesion, cell differentiation, de and recellularization, esophagus, stem cells, tissue engineering, tissue, recellularization, transplantation, quantification, Biochemistry & Molecular Biology
Subject categories Surgery

Abstract

Decellularization of esophagus was studied using three different protocols. The sodium deoxycholate/DNase-I (SDC/DNase-I) method was the most successful as evidenced by histology and DNA quantification of the acellular scaffolds. Acellular scaffolds were further analyzed and compared with native tissue by histology, quantitative analysis of DNA, and extracellular matrix (ECM) proteins. Histologically, the SDC/DNase-I protocol effectively produced scaffold with preserved structural architecture similar to native tissue architecture devoid of any cell nucleus. ECM proteins, such as collagen, elastin, and glycosaminoglycans were present even after detergent-enzymatic decellularization. Immunohistochemical analysis of acellular scaffold showed weak expression of Gal 1, 3 Gal epitope compared with native tissue. For performing recellularization, human amnion-derived mesenchymal stem cells (MSCs) and epithelial cells were seeded onto acellular esophagus in a perfusion-rotation bioreactor. In recellularized esophagus, immunohistochemistry showed infiltration of MSCs from adventitia into the muscularis externa and differentiation of MSCs into the smooth muscle actin and few endothelial cells (CD31). Our study demonstrates successful preparation and characterization of a decellularized esophagus with reduced load of Gal 1, 3 Gal epitope with preserved architecture and ECM proteins similar to native tissue. Upon subsequent recellularization, xenogeneic acellular esophagus also supported stem cell growth and partial differentiation of stem cells. Hence, the current study offers the hope for preparing a tissue-engineered esophagus in vitro which can be transplanted further into pigs for further in vivo evaluation.

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