In vitro exposure of pig neonatal isletlike cell clusters to human blood

• Published in: Xenotransplantation (Københaven) Vol. 22; no. 4; pp. 317 - 324
• Main Authors: Nagaraju, Santosh, Bertera, Suzanne, Tanaka, Takayuki, Hara, Hidetaka, Rayat, Gina R., Wijkstrom, Martin, Ayares, David, Trucco, Massimo, Cooper, David K. C., Bottino, Rita
• Format: Journal Article
• Language: English
• Published: Denmark Blackwell Publishing Ltd 01.07.2015
• Subjects: Animals; Animals, Genetically Modified; Animals, Newborn; Antigens, CD - genetics; Antigens, CD - immunology; Apyrase - genetics; Apyrase - immunology; Blood - immunology; Blood Coagulation; Complement Activation; diabetes mellitus; Diabetes Mellitus - therapy; Galactosyltransferases - deficiency; Galactosyltransferases - genetics; Galactosyltransferases - immunology; Gene Knockout Techniques; genetically engineered; Humans; In Vitro Techniques; instant blood-mediated inflammatory reaction; islets; Islets of Langerhans Transplantation - adverse effects; Islets of Langerhans Transplantation - immunology; Islets of Langerhans Transplantation - pathology; Membrane Cofactor Protein - genetics; Membrane Cofactor Protein - immunology; neonatal; pigs; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Sus scrofa; Transplantation, Heterologous - adverse effects; Transplantation, Heterologous - methods; xenotransplantation; pigs; genetically engineered; islets; neonatal; instant blood-mediated inflammatory reaction; diabetes mellitus; xenotransplantation
• ISSN: 0908-665X; 1399-3089; 1399-3089
• DOI: 10.1111/xen.12178

Background Pig islet grafts have been successful in treating diabetes in animal models. One remaining question is whether neonatal pig isletlike cell clusters (NICC) are resistant to the early loss of islets from the instant blood‐mediated inflammatory reaction (IBMIR). Methods Neonatal isletlike cell clusters were harvested from three groups of piglets—(i) wild‐type (genetically unmodified), (ii) α1,3‐galactosyltransferase gene‐knockout (GTKO)/CD46, and (iii) GTKO/CD46/CD39. NICC samples were mixed with human blood in vitro, and the following measurements were made—antibody binding; complement activation; speed of islet‐induced coagulation; C‐peptide; glutamic acid decarboxylase (GAD65) release; viability. Results Time to coagulation and viability were both reduced in all groups compared to freshly drawn non‐anticoagulated human blood and autologous combinations, respectively. Antibody binding to the NICC occurred in all groups. Conclusions Neonatal isletlike cell clusters were subject to humoral injury with no difference associated to their genetic characteristics.