GSM4116267: Single cell mRNA-seq_PC9_DMSO_invivo; Homo sapiens; RNA-Seq
Single cell isolation form tumors: Fresh tumor specimens (n=3) were pooled and minced in a 15 mL conical tube with media (DMEM + 10% FBS), penicillin–streptomycin (Fisher Scientific), 100 U/mL collagenase type IV (Life Technologies) and 2.5 mg/mL DNAse I (Sigma Aldrich), then incubated for 45min at 37°C.  Single cell suspensions were isolated by straining through a 40μm filters.  Cells were incubated with  Zombie Green™ Fixable Viability Kit (BioLegend), blocked with Human TruStain FcX™ (BioLegend), and stained with human anti-EpCAM (clone 9C4).  Viable EpCAM+ tumor cells were isolated via FACS Melody instrument (BD Biosciences). Cells were loaded onto a 10X Chromium instrument (10X Genomics) per the manufacturer's instructions. Single-cell RNA libraries were generated using the Single Cell 3' Reagent Kit (10X Genomics) per user guide.  Quality control of the completed libraries was performed using Bioanalyzer High Sensitivity DNA Kit (Agilent)
GEO Accession
GSM4116267