GSM6333723: Fibro2_DMSO; Mus musculus; RNA-Seq
For fibroblast isolation, cells were isolated from Balb/cAnN mice orthotopically transplanted with 4T1 tumours pre-treated with DMSO or GSK. Single-cell suspensions were incubated with mouse FcR Blocking Reagent and subsequently with the following APC labelled antibodies – CD45, CD31, CD326 and hCD2. Cells were washed and incubated with magnetic anti-APC MicroBeads (Miltenyi, 130-090-855). Magnetically labelled immune, endothelial, epithelial and cancer cells were isolated using LD columns, washed thrice with 3ml of MACS and the unlabelled flow-through cell fraction was the mesenchymal cells that we will refer to as fibroblasts hereafter. To increase the purity of the cells, the eluted fraction was applied to a second round of separation with a new LD column.