SRS243 - Modelling ABO blood group mediated hyperacute rejection using precision cut kidney slices from transplant-declined human organs

Abstract Background Precision cut kidney slices (PCKS) are a valuable model to bridge the gap between cellular and in vivo research. They offer an experimental platform that maintains many features of an intact organ, including its heterogeneous cellular architecture and some of its function whilst enabling higher throughput and greater relevance than animal models. Here, we describe the development of a novel PCKS model of complement activation in an ABO blood group incompatible environment. Methods The Alabama R&D tissue slicer was used to produce PCKS from human blood group A kidneys declined for transplantation and offered for research. Tissue integrity was investigated using lactate dehydrogenase (LDH) release assays, and formalin-fixed paraffin embedded tissue sections stained with hematoxylin and eosin. PCKS were then incubated with complement active human blood group O serum for up to 4 h. Complement activation at 2 and 4 h was investigated using enzyme-linked immunosorbent assays, microscopy, and viability assays. Results PCKS tissue viability was maintained over the 4-hour period. There were no significant differences in LDH release over the time period or in nuclei counts compared to pre samples. A marked increase in medium C5a levels was observed in PCKS incubated with incompatible human blood group O serum, indicating complement activation. Further, increased levels of LDH were identified in the medium of PCKS exposed to an ABO incompatible environment, indicating loss of membrane integrity. Conclusions This new PCKS model provides a valuable platform to investigate hyperacute rejection and possible treatments in a highly relevant experimental model.