Improving KIdney Transplantation with Cellular Therapy Study

This observational, cohort study will be conducted at Westmead Hospital * 3 visits over 24-months per participant. * each consenting participant will donate 20 - 50ml of peripheral blood per visit. * recruit n = 20 per study group (control, CKD, dialysis, and transplant). Aims include: 1. Compare generation of therapeutic grade tolerogenic dendritic cells (TolDC) from peripheral blood mononuclear cells derived from controls (healthy) and patients with kidney disease (CKD, dialysis, or transplantation). 2. Perform testing of TolDC phenotype, tolerogenic capacity and yield. 3. Generate hypothesis for different mechanisms of TolDC function not yet described for further characterisation to support future research Research plan: * Peripheral blood mononuclear cells (PBMC) will be isolated from blood by Ficoll gradient and ultrapure magnetic bead sorting. * PBMC will be tested for purity (other unwanted immune cells) and microbiological contaminants before added to specialized DC media to generate TolDC. * TolDC will have in depth functional characterization including flow cytometry, cytokine expression, genomic sequencing, and in-vitro testing protocols. * TolDCs will be co-cultured with human renal cells to test protective capabilities across different AKI stimuli/insults. This allows selection of ideal PBMC donors to generate functionally desirable TolDC for subsequent use in i-KITCaT studies.