GSM7728595: Pericardial fluid; Homo sapiens; RNA-Seq
Heart tissue, spleen tissue, pericardial fluid, and blood in the heart were obtained. The heart tissue was dissociated in DMEM containing Collagenase, Type 1 (2 mg/ml, Thermo Fisher Scientific, MA, USA) and DNase1 (100µg/ml, Roche, Basel, Switzerland) at 37 °C for 60 minutes using MACSmix Tube Rotator (Miltenyi Biotec, North Rhine-Westphalia, Germany). 150ml of pericardial fluid and 30ml of blood in the heart were collected (with heparin for the blood). To remove normal-density neutrophils, the cells isolated from pericardial fluid and heart blood were layered over lymphocyte separation solution (Nacalai Tesque, Kyoto, Japan). The sample was centrifuged at 1200 G for 10 minutes, and mononuclear cells from heart tissue were harvested. Mononuclear cells from pericardial fluid and blood were harvested and used immediately for analysis. CD45 positive cells from the heart tissue were selected using EasySep Release Human CD45 Positive Selection Kit (Stemcell Technologies, British Columbia, Canada) and used immediately. Libraries were generated using the Chromium Single Cell 5' Reagent kit on Chromium Single-Cell Controller (10x Genomics, CA, USA) according to the manufacturers' instructions.