title: GSM3464138: 2016_DB_DF_06; Homo sapiens; RNA-Seq
library_construction_protocol: Human dermal fibroblasts were isolated from systemic sclerosis patients (fibrotic fibroblasts) and age- and sex-matched healthy volunteers (resting fibroblasts). After enzymatic digestion of the skin biopsies with collagenase type II (Merck, Darmstadt, 676 Germany) for 3h at 37°C, the digested tissues were filtered and centrifuged at 1400 rpm for 5 min. The pellet was resuspended in DMEM/F-12 medium containing 10% heat inactivated fetal bovine serum (FBS), 25 mM HEPES, 100 U/ml penicillin, 100 μg/ml streptomycin, 2 mM L-glutamine, 2.5 μg/ml amphotericin B (Thermo Fisher Scientific, Dreieich, Germany) and 0.2 mM ascorbic acid (Sigma-Aldrich, Steinheim, Germany). Synovial fibroblasts (inflammatory fibroblasts) were isolated from inflamed joints of patients with rheumatoid arthritis. Fibroblasts were cultured for 3 passages and then quality checked for a pure fibroblast population before using them in experiments. Fibroblasts were negative for CD31, CD45, CD326 (EpCAM) and KRT14 and positive for collagen-1, PDGFRα and vimentin. RNA libraries were prepared for sequencing using Illumina stranded mRNA protocols
attributes.tag: GEO Accession
attributes.value: GSM3464138