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

Xenotransplantation. 2015 Jul-Aug;22(4):317-24. doi: 10.1111/xen.12178. Epub 2015 Jul 14.

Abstract

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.

Keywords: diabetes mellitus; genetically engineered; instant blood-mediated inflammatory reaction; islets; neonatal; pigs; xenotransplantation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Animals, Newborn
  • Antigens, CD / genetics
  • Antigens, CD / immunology
  • Apyrase / genetics
  • Apyrase / immunology
  • Blood / immunology*
  • Blood Coagulation
  • Complement Activation
  • Diabetes Mellitus / therapy
  • Galactosyltransferases / deficiency
  • Galactosyltransferases / genetics
  • Galactosyltransferases / immunology
  • Gene Knockout Techniques
  • Humans
  • In Vitro Techniques
  • Islets of Langerhans Transplantation / adverse effects
  • Islets of Langerhans Transplantation / immunology*
  • Islets of Langerhans Transplantation / pathology
  • Membrane Cofactor Protein / genetics
  • Membrane Cofactor Protein / immunology
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Sus scrofa
  • Transplantation, Heterologous / adverse effects
  • Transplantation, Heterologous / methods*

Substances

  • Antigens, CD
  • CD46 protein, human
  • Membrane Cofactor Protein
  • Recombinant Proteins
  • Galactosyltransferases
  • alpha-1,3-galactosyltransferase 1, porcine
  • Apyrase
  • CD39 antigen