GSM5751918: scRNA-seq, human, resistant; Homo sapiens; RNA-Seq
Preparation: collect the fresh tumor tissue from MIBC patients and wash the samples with PBS for 3 times. After the tissue preparation, collagen I (0.5mgml-1) and collagen IV (1mgml-1) was combined to digest the samples with 2 hours in 37℃. The 400g centrifuge was used with 6 minutes. After discarding the supernatant, 10 ml trypsin was used to resuspend with 15 minutes in 37℃. The 10% FBS was added to medium, and filter the mixture through 100μm. Then 400g centrifuge was used. After discarding the supernatant, 4ml 1XACK was used to resuspend the cells on the ice with 3 minutes. And 200 μl 0.04%BSA-PBS was used to resuspend cells. After quantifying the cell number, adjusting the concentration to the 800-1200 cells/μl and ensuring the cell activity > 85%. Follwed by v2 protocol of chemistry of the 3' scRNA-seq 10X genomics platform