Illumina NovaSeq 6000 paired end sequencing; Primed to resolve: A single cell atlas of the shoulder capsule reveals a cellular basis for resolving inflammatory fibrosis (Bulk RNA-seq of capsular fibroblasts co-cultured with MDMs)
Primed to resolve: A single cell atlas of the shoulder capsule reveals a cellular basis for resolving inflammatory fibrosis (Bulk RNA-seq of capsular fibroblasts co-cultured with MDMs)
Capsular fibroblasts were isolated from frozen shoulder patients and were co-cultured with monocyte-derived macrophages(MDM), which were isolated from blood cones and treated with M-CSF for 5 days. Capsule-derived fibroblasts isolated from frozen shoulder patients were added to MerTKhigh or MerTKlow incubations at a density of 200,000 cells/well. Fibroblasts and MDMs were co-cultured for 48 hrs in complete media containing 3% FCS prior to harvest and sorting. RNA was extracted using RNEasy microprep kit (Qiagen) according to the manufacturers protocol.  Double stranded cDNA library construction was performed using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (E7760L) with custom adapters and dual indexed barcodes.