Isolated cells were counted and suspended in 0.05%BSA (Roche) in PSB. Cells were stained with mouse CD45 PeCy7 and human CD45 FITC (BD Biosciences) following suppliers’ instructions and analysed by flow cytomentry (Attune, Thermo)
Fixed tissues from the efficacy study mice were processed for immunochistochemistry by Cellullar Pathology, RVI Newcastle Hospitals NHS trust using standard methods as follows. Briefly, bones were decalcified using EDTA. Tissues were infiltrated with and embedded in parrafin wax. Sections on slides were staining with either, hematoxylin and eosin (H&E) or, human CD19 antibody using a Ventana BenchMark Ultra (Ventana, Roche), and Universal DAB Detection kit (Ultraview) to produce a brown chromogen at the site of human CD19. Slides were scanned using an Aperio ScanScope (Leica) and images analysised using Leica eSlide manager software.
Fixed tissues from the efficacy study mice were processed for immunochistochemistry by Cellullar Pathology, RVI Newcastle Hospitals NHS trust using standard methods as follows. Briefly, bones were decalcified using EDTA. Tissues were infiltrated with and embedded in parrafin wax. Sections on slides were staining with either, hematoxylin and eosin (H&E) or, human CD19 antibody using a Ventana BenchMark Ultra (Ventana, Roche), and Universal DAB Detection kit (Ultraview) to produce a brown chromogen at the site of human CD19. Slides were scanned using an Aperio ScanScope (Leica) and images analysised using Leica eSlide manager software.