GSM3184687
GSM3184687: single_cell_RNAseq Young10X2; Mus musculus; RNA-Seq
Animals were sacrificed by cervical dislocation and their brains immediately placed on ice. The lateral SVZ was microdissected as a whole mount as previously described. Pooled SVZ tissue from 4 mice of 2 and 8 mice of 22 months of age were pulled for each library. Tissue was digested with trypsin and DNAse according to the Neural Tissue Dissociation kit in a Gentle MACS dissociator (Miltenyi) to maximize cell viability and recovery. Cells were stained for 20 minutes on ice in DPBS (without Ca and Mg) supplemented with 10% fetal calf serum. Combinations of the following antibodies were used:  Ter119 APC-Cy7 (Biolegend, 1:100), O4 APC-Cy7 (Miltenyi, 1:50), CD45 APC-Cy7 (BD, 1:200) , GLAST(ACSA-1)-PE (Miltenyi, 1:20). Sytox Blue (Life Technologies, 1/1000) was added to the cell suspension prior to sorting. Stained cell suspensions were sorted in a FACS AriaII (BD) using a 100um nozzle at 20psi. Cells were collected in ice-cold Neurobasal medium supplemented with L-Glutamine, B27. To ensure that there is enough starting material for the 10x platform, we did a negative selection for O4, Ter119 and CD45 and we positively selected for GLAST. 8000 FACS events underwent the downstream process according to the manufacturer's details. Libraries were prepared according to Chromium™ Single Cell 3' Reagent Kit.
GEO
GSM3184687
GEO Accession
GSM3184687