GSM5293860: D1; Homo sapiens; RNA-Seq
Human tissue was obtained from healthy donors and patients. Place a sterile RNase-free petri dish containing 1 ml Ca2+ and Mg2+ free Phosphate Buffer Saline (PBS) , the tissue in the protection solution  (Miltenyi, Bergisch Gladbach, Germany, 130-100-008) was transferred into the petri dish and scraped the fluff with a scalpel. The tissue was transfered into a 2 ml centrifuge tube and digested at 37°C for 7 min in 0.25% (W/V) of trypsin. After Aspirated the trypsin solution, the cell suspension was washed twice with preheated PBS  (Ca2+, Mg2+ free) and aspirated the cleaning solution. Digestion of the cell suspension was at 37°C for 20 minutes with 1mg/ml collagenase I, 1mg/ml dispase and 0.05mg/ml DNase I and observed under microscope to obtain cells and the cell suspension was filtered by passing through a 40 μm cell sieve and centrifuged at 300g for 5 min at 4°C after digestion. The cell precipitation was resuspended in PBS (Ca2+, Mg2+ free) and lysed the red blood cells by Lysing Buffer (Gibco ACK, A1049201). The suspension was centrifuged at 300g for 5 min at 4°C to pellet cells and removed dead cells and cell debris by washing twice with PBS (Ca2+, Mg2+ free). Microscopic examination was performed to confirm the overall cell viability.