GSM8679267: Animal R235, T Cells, Liver, GEX; Macaca mulatta; RNA-Seq
MHC-haploidentical HCT and autologous HCT underwent pre-transplant conditioning with 1040 cGy total body irradiation prior to transplantation with an apheresis-derived peripheral blood stem cell product. Transplant recipients were not given immunosuppression. Animals were euthanized when they met a pre-set experimental endpoint on day 8 or prespecified clinical endpoints. The lungs and liver were cut into small pieces and digested with Dnase I and type I collagenase at 37°C for 1 hour, and passed through a metal strainer, then 100-, 70-, and 40-μm nylon strainers. The resulting cell suspension was layered into a 70% and 30% Percoll gradient and the leukocytes were collected at the interphase between the layers. The cells were washed and immediately analyzed or cryopreserved in 10% DMSO in FBS and stored in liquid nitrogen. Cells were processed for single cell sequencing using the Next GEM 5' v2 Gel Beads and either i7 Multiplex Plate (Single Index) or Single Cell 5' v2 Sample Index Plate TT (Dual Index). All products used in the preparation of the samples are available from 10x Genomics, except the NHP TCR specific custom designed primers that were developed as previously described (DOI 10.3389/fimmu.2021.804932). These NHP TCR specific custom primer set were substituted for the provided primers in the T Cell Mix 1 and 2 premixed primers.