Inflammation Induced by Cold Ischemia: A New Fast and Simple In Vivo Method to Evaluate Potential Therapeutic Tools

NELLA AMBROSI, DIEGO GUERRIERI, CAROLINA BOUSO, GABRIELA SALAMONE, JOSEFINA MENDIBERRI, CLAUDIO INCARDONA, EDUARDO CHULUYAN, DOMINGO CASADEI

Congreso; World Transplant Congress; 2014

Background: a prolonged cold ischemic (CI) time in kidney transplants has been associated with a high rate of delayed graft function, acute rejection, and reduced graft survival. The aim of this work was to develop a simple and rapid method that allows us to evaluate, in vivo, potential therapeutic tools, which may reduce the inflammatory process caused by the prolonged CI time in renal tissue. Materials and Methods: Kidney explants (Exp) derived from BALB/c wild type mice (WT) were incubated ex vivo at 4 ° C for 18 h (CI-Exp) in different preservation solutions. At the end of the incubation period, explants were implanted subcutaneously (s.c.) in WT or transgenic mice expressing green fluorescent protein (GFP). As control group and, at the same time, these animals also received explants that do not suffered CI. After 24 hours, mice were sacrificed and the explants were recovered and processed, in order to examine the recruitment of neutrophils. The recruitment of these cells was examined by measuring myeloperoxidase (MPO), as an indirect marker of neutrophils migration, and by a direct counting of the Gr1-Phe+ and GFP+ cells with a flow cytometer. Results: explants that were incubated at 4ºC for 18 h in Ringer Lactate (RL), recruited more neutrophils than control explants (Control: 0.3 ± 0.06; CI-explants: 0.6 ± 0.08, p