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.
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.



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).



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.



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.
1 Department of Medicine, University of Vermont College of Medicine , Burlington, Vermont.
Joseph
J
Platz
1 Department of Medicine, University of Vermont College of Medicine , Burlington, Vermont.
Nicholas R
NR
Bonenfant
1 Department of Medicine, University of Vermont College of Medicine , Burlington, Vermont.
Franziska E
FE
Uhl
1 Department of Medicine, University of Vermont College of Medicine , Burlington, Vermont.
Amy L
AL
Coffey
1 Department of Medicine, University of Vermont College of Medicine , Burlington, Vermont.
Tristan
T
McKnight
1 Department of Medicine, University of Vermont College of Medicine , Burlington, Vermont.
Charles
C
Parsons
1 Department of Medicine, University of Vermont College of Medicine , Burlington, Vermont.
Dino
D
Sokocevic
1 Department of Medicine, University of Vermont College of Medicine , Burlington, Vermont.
Zachary D
ZD
Borg
2 Department of Biology and VGN Proteomics Facility, University of Vermont College of Arts and Sciences , Burlington, Vermont.
Ying-Wai
YW
Lam
2 Department of Biology and VGN Proteomics Facility, University of Vermont College of Arts and Sciences , Burlington, Vermont.
Bin
B
Deng
2 Department of Biology and VGN Proteomics Facility, University of Vermont College of Arts and Sciences , Burlington, Vermont.
Julia G
JG
Fields
3 Biostatistics Unit, University of Vermont College of Medicine , Burlington, Vermont.
Michael
M
DeSarno
4 Department of Biomedical Sciences, University of Cagliari , Cagliari, Italy .
Roberto
R
Loi
5 Department of Clinical Sciences, Tufts University , Cummings School of Veterinary Medicine, North Grafton, Massachusetts.
Andrew M
AM
Hoffman
6 Revivicor, Inc. , Blacksburg, Virginia.
John
J
Bianchi
7 Exemplar Genetics, Sioux Center , Iowa.
Brian
B
Dacken
8 United Therapeutics Corp., Research Triangle Park , Durham, North Carolina.
Thomas
T
Petersen
1 Department of Medicine, University of Vermont College of Medicine , Burlington, Vermont.|9 Comprehensive Pneumonology Center, Helmholtz Center Munich, Ludwig Maximilians University Munich , Munich, Germany .
Darcy E
DE
Wagner
1 Department of Medicine, University of Vermont College of Medicine , Burlington, Vermont.
Daniel J
DJ
Weiss
D000818:Animals; D030801:Animals, Genetically Modified; D049109:Cell Proliferation; D004847:Epithelial Cells / Q000166:cytology* / Q000201:enzymology; D005109:Extracellular Matrix / Q000201:enzymology; D005347:Fibroblasts / Q000166:cytology* / Q000201:enzymology; D005700:Galactosyltransferases / Q000502:physiology*; D006801:Humans; D008168:Lung / Q000166:cytology* / Q000201:enzymology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000201:enzymology; D012038:Regeneration / Q000502:physiology*; D013552:Swine
D005700:Galactosyltransferases; C047006:N-acetyllactosaminide alpha-1,3-galactosyltransferase