Science and Research

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

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 (alpha-gal) activity, that could conceivably prevent effective use. Use of lungs from alpha-gal knock out (alpha-gal KO) pigs presents a potential alternative and thus comparative de- and recellularization of wild-type and alpha-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 alpha-gal KO pig lungs, including retention of alpha-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 alpha-gal KO lungs. CONCLUSION: These proof of concept studies demonstrate that decellularized wild-type and alpha-gal KO pig lungs can be comparably decellularized and comparably support initial growth of human lung cells, despite some differences in retained proteins. alpha-Gal KO pig lungs are a suitable platform for further studies of xenogeneic lung regeneration.

  • Platz, J.
  • Bonenfant, N. R.
  • Uhl, F. E.
  • Coffey, A. L.
  • McKnight, T.
  • Parsons, C.
  • Sokocevic, D.
  • Borg, Z. D.
  • Lam, Y. W.
  • Deng, B.
  • Fields, J. G.
  • DeSarno, M.
  • Loi, R.
  • Hoffman, A. M.
  • Bianchi, J.
  • Dacken, B.
  • Petersen, T.
  • Wagner, D. E.
  • Weiss, D. J.

Keywords

  • Animals
  • Animals, Genetically Modified
  • Cell Proliferation
  • Epithelial Cells/*cytology/enzymology
  • Extracellular Matrix/enzymology
  • Fibroblasts/*cytology/enzymology
  • Galactosyltransferases/*physiology
  • Humans
  • Lung/*cytology/enzymology
  • Mesenchymal Stromal Cells/*cytology/enzymology
  • Regeneration/*physiology
  • Swine
Publication details
DOI: 10.1089/ten.TEC.2016.0109
Journal: Tissue Eng Part C Methods
Pages: 725-39 
Number: 8
Work Type: Original
Location: CPC-M
Disease Area: ROR
Partner / Member: HMGU
Access-Number: 27310581
See publication on PubMed

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