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

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 detergentenzymatic 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.

Renal Regeneration: The Role of Extracellular Matrix and Current ECM‐Based Tissue Engineered Strategies

“…[101] Adult stem cells from kidney tissue obtained from clonal nephrospheres have also demonstrated to spontaneously differentiate into parenchymal and endothelial cell populations in kidney ECM scaffolds by 30 days of culture time, demonstrating the time-dependent role of ECM in cellular differentiation. [102] The engraftment and differentiation of different adult stem cells was already demonstrated in decellularized matrices of vascular grafts, [103] human cornea scaffolds, [104] pig esophagus, [105] in an aortic patch, [106] blood vessels, [107] myocardium scaffolds, [108,109] cardiac ECM, [110] small intestine, [111] lung, [112] and tracheal allografts. [113] As decellularized matrices have shown to provide adequate stimulus for uncommitted and committed cells to adhere, proliferate and differentiate into the specific organ-lineages, it was noticed that these matrices do not only preserve the specificity of tissues, but also the stage of development of the organ.…”

“…[ 101 ] Adult stem cells from kidney tissue obtained from clonal nephrospheres have also demonstrated to spontaneously differentiate into parenchymal and endothelial cell populations in kidney ECM scaffolds by 30 days of culture time, demonstrating the time‐dependent role of ECM in cellular differentiation. [ 102 ] The engraftment and differentiation of different adult stem cells was already demonstrated in decellularized matrices of vascular grafts, [ 103 ] human cornea scaffolds, [ 104 ] pig esophagus, [ 105 ] in an aortic patch, [ 106 ] blood vessels, [ 107 ] myocardium scaffolds, [ 108,109 ] cardiac ECM, [ 110 ] small intestine, [ 111 ] lung, [ 112 ] and tracheal allografts. [ 113 ]…”

Biofabrication of an Esophageal Tissue Construct from a Polymer Blend Using 3D Extrusion‐Based Printing

“…[ 2 ] Unfortunately, the resection of the esophagus can lead to postoperative complications and surgical‐associated morbidities and mortality. [ 3,4 ] The use of decellularized esophageal transplants is considered a favorable alternative [ 5,6 ] but has a number of drawbacks, e.g., donor shortage, inadequate size, poor mechanical performance, immunogenicity, rapid degradation, and the need of a stent support, among others. [ 6,7 ]…”

Genipin-crosslinked decellularized scaffold induces regeneration of defective rat kidneys

“…Compared with other crosslinking agents, such as glutaraldehyde, genipin has many advantages, such as noncytotoxicity. 27 In addition to its nontoxic characteristics to tissue cells, genipin used as a crosslinking agent in tissue engineering bioscaffold materials has been proven to improve the performance of decellularized biological scaffolds in multiple organs, such as decellularized trachea, 28 esophagus 29 and even in complex organs such as the uterus. 30 The biological properties of the stent are improved after crosslinking with genipin.…”