Illumina NovaSeq 6000 sequencing; Fibroblastic reticular cell lineage convergence in Peyer's patches governs intestinal homeostasis
Fibroblastic reticular cell lineage convergence in Peyer's patches governs intestinal homeostasis
Peyer's patches were harvested and washed with cold PBS. The tissue was digested using an enzyme mix comprised of RPMI containing 10% fetal bovine serum, 0.8 mg/ml Dispase, 0.1 mg/ml Collagenese P (Roche) and 0.1 mg/ml DNase I (Sigma) for 45 min at 37 °C. Cells were washed with cold PBS and suspended in FACS buffer (PBS supplemented with 0.5% bovine serum albumin (BSA) and 5 mM EDTA in PBS). Hematopoietic cells were depleted using anti-CD45 microbeads and magnetic based separation (Miltenyi). Remaining cells were stained with labelled antibodies and FRCs were sorted by FACS sorting. The cDNA library generation was performed following the established commercial protocol for Chromium Single Cell 3' Reagent Kit (v2 Chemistry or v3 Chemistry)
Institute of Immunobiology, Kantonsspital St. Gallen, St. Gallen, Switzerland
E-MTAB-9162:Sample 6_p