GSM2124704: SUM149PT_co-culture; Homo sapiens; OTHER
DsRED labeled carcinoma cell lines and GFP expressing primary CAFs were cultured either separately or in co-cultures for 17 hours. Colonies were released from Matrigel by 30 min incubation in PBS/EDTA, at which point separately cultured cells were combined to control for cross-contamination. Cell mixtures were washed with PBS, trypsinized for 7 min, quenched with serum containing media, washed and spun down. Epithelial cells were selected using Epithelial Enrich dynabeads (Life Technologies) following manufacturer’s protocol. Microscopic examination of captured cells revealed no detectable CAF contamination. From each sample 5E5 cells were used to prepare SAGE-Seq (serial analysis of gene expression combined with the Illumina/Solexa 1G platform) libraries according to a detailed protocol available at http://polyaklab.dfci.harvard.edu/index.php/science/protocols
GEO Accession
GSM2124704