Age-related thymic involution modifies epithelial composition, function and T cell receptor repertoire

We aimed to investigate changes in T cell receptor (TCR) sequence repertoire over different ages in mouse thymocytes after negative selection. We used FACS to isolate M2 thymocytes from mice aged 1, 4, 16, 32 or 52 weeks old. RNA was extracted from 15,000 cells per sample. The Smarter Mouse TCR a/b Profiling Kit (Takara) was used to generate TCR-seq libraries and these were sequenced on a MiSeq (300 PE reads).