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Comparative Decellularization and Recellularization of Wild-Type and Alpha 1,3 Galactosyltransferase Knockout Pig Lungs : A Model for Ex Vivo Xenogeneic Lung Bioengineering and Transplantation

Platz, Joseph; Bonenfant, Nicholas R.; Uhl, Franziska E.; Coffey, Amy L.; McKnight, Tristan; Parsons, Charles; Sokocevic, Dino; Borg, Zachary D.; Lam, Ying Wai and Deng, Bin, et al. (2016) In Tissue Engineering - Part C: Methods 22(8). p.725-739
Abstract

Background: A novel potential approach for lung transplantation could be to utilize xenogeneic decellularized pig lung scaffolds that are recellularized with human lung cells. However, pig tissues express several immunogenic proteins, notably galactosylated cell surface glycoproteins resulting from alpha 1,3 galactosyltransferase (α-gal) activity, that could conceivably prevent effective use. Use of lungs from α-gal knock out (α-gal KO) pigs presents a potential alternative and thus comparative de- and recellularization of wild-type and α-gal KO pig lungs was assessed. Methods: Decellularized lungs were compared by histologic, immunohistochemical, and mass spectrometric techniques. Recellularization was assessed following compartmental... (More)

Background: A novel potential approach for lung transplantation could be to utilize xenogeneic decellularized pig lung scaffolds that are recellularized with human lung cells. However, pig tissues express several immunogenic proteins, notably galactosylated cell surface glycoproteins resulting from alpha 1,3 galactosyltransferase (α-gal) activity, that could conceivably prevent effective use. Use of lungs from α-gal knock out (α-gal KO) pigs presents a potential alternative and thus comparative de- and recellularization of wild-type and α-gal KO pig lungs was assessed. Methods: Decellularized lungs were compared by histologic, immunohistochemical, and mass spectrometric techniques. Recellularization was assessed following compartmental inoculation of human lung bronchial epithelial cells, human lung fibroblasts, human bone marrow-derived mesenchymal stromal cells (all via airway inoculation), and human pulmonary vascular endothelial cells (CBF) (vascular inoculation). Results: No obvious differences in histologic structure was observed but an approximate 25% difference in retention of residual proteins was determined between decellularized wild-type and α-gal KO pig lungs, including retention of α-galactosylated epitopes in acellular wild-type pig lungs. However, robust initial recellularization and subsequent growth and proliferation was observed for all cell types with no obvious differences between cells seeded into wild-type versus α-gal KO lungs. Conclusion: These proof of concept studies demonstrate that decellularized wild-type and α-gal KO pig lungs can be comparably decellularized and comparably support initial growth of human lung cells, despite some differences in retained proteins. α-Gal KO pig lungs are a suitable platform for further studies of xenogeneic lung regeneration.

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published
in
Tissue Engineering - Part C: Methods
volume
22
issue
8
pages
725 - 739
publisher
Mary Ann Liebert Inc.
external identifiers
  • scopus:84983762681
ISSN
1937-3384
DOI
10.1089/ten.tec.2016.0109
language
English
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no
id
11f38a2d-43a1-4149-b5d0-e93a51bc81e8
date added to LUP
2017-08-15 14:45:41
date last changed
2017-08-22 16:12:09
@article{11f38a2d-43a1-4149-b5d0-e93a51bc81e8,
  abstract     = {<p>Background: A novel potential approach for lung transplantation could be to utilize xenogeneic decellularized pig lung scaffolds that are recellularized with human lung cells. However, pig tissues express several immunogenic proteins, notably galactosylated cell surface glycoproteins resulting from alpha 1,3 galactosyltransferase (α-gal) activity, that could conceivably prevent effective use. Use of lungs from α-gal knock out (α-gal KO) pigs presents a potential alternative and thus comparative de- and recellularization of wild-type and α-gal KO pig lungs was assessed. Methods: Decellularized lungs were compared by histologic, immunohistochemical, and mass spectrometric techniques. Recellularization was assessed following compartmental inoculation of human lung bronchial epithelial cells, human lung fibroblasts, human bone marrow-derived mesenchymal stromal cells (all via airway inoculation), and human pulmonary vascular endothelial cells (CBF) (vascular inoculation). Results: No obvious differences in histologic structure was observed but an approximate 25% difference in retention of residual proteins was determined between decellularized wild-type and α-gal KO pig lungs, including retention of α-galactosylated epitopes in acellular wild-type pig lungs. However, robust initial recellularization and subsequent growth and proliferation was observed for all cell types with no obvious differences between cells seeded into wild-type versus α-gal KO lungs. Conclusion: These proof of concept studies demonstrate that decellularized wild-type and α-gal KO pig lungs can be comparably decellularized and comparably support initial growth of human lung cells, despite some differences in retained proteins. α-Gal KO pig lungs are a suitable platform for further studies of xenogeneic lung regeneration.</p>},
  author       = {Platz, Joseph and Bonenfant, Nicholas R. and Uhl, Franziska E. and Coffey, Amy L. and McKnight, Tristan and Parsons, Charles and Sokocevic, Dino and Borg, Zachary D. and Lam, Ying Wai and Deng, Bin and Fields, Julia G. and DeSarno, Michael J and Loi, Roberto and Hoffman, Andrew M. and Bianchi, John and Dacken, Brian and Petersen, Thomas N and Wagner, Darcy E. and Weiss, Daniel J.},
  issn         = {1937-3384},
  language     = {eng},
  month        = {08},
  number       = {8},
  pages        = {725--739},
  publisher    = {Mary Ann Liebert Inc.},
  series       = {Tissue Engineering - Part C: Methods},
  title        = {Comparative Decellularization and Recellularization of Wild-Type and Alpha 1,3 Galactosyltransferase Knockout Pig Lungs : A Model for Ex Vivo Xenogeneic Lung Bioengineering and Transplantation},
  url          = {http://dx.doi.org/10.1089/ten.tec.2016.0109},
  volume       = {22},
  year         = {2016},
}