A, Viability by trypan blue exclusion of colorectal cells digested from biopsies either fresh, cryopreserved as biopsies, or cryopreserved after digestion (n = 18 tissue donors). B, Live cell yield by trypan blue exclusion. C, Recovery of live cells after cryopreservation relative to fresh samples from the same donor. D, Background-subtracted cytokine production from colorectal T cells after stimulation with cytomegalovirus, Epstein-Barr virus and influenza virus peptides (CEF); HIV Gag peptides (Gag); phorbol 12-myristate 13-acetate and ionomycin (PMA/Iono); or staphylococcal enterotoxin B (SEB). Small gray symbols indicate individual samples, with gray lines connecting samples from the same tissue donors. Colored symbols show the mean across all samples from HIV+ (orange) and HIV- (green) donors, with vertical lines showing the 95% confidence interval of the mean. E, Polyfunctionality of colorectal responses to stimulation. Each sub-bar corresponds to the percent of cells with the indicated number of functions, averaged across the donors. Total bar height indicates the total percent of cells with one or more functions.