GSM8678773: Mouse retina, 12mo, Rep 3; Mus musculus; RNA-Seq
2 non-injured retinae were collected from C57BL/6J mice per condition. Retinal tissue was digested using the papain dissociation system (Worthington) following the manufacturer's protocol. Cells were passed through a 30 µm filter (Miltenyi) to obtain a single-cell suspension. Following this different magnetic activated cell sorting methods (MACS®) enrichment/depletion methods were used on the cell pool collected. ACSA-2 and Thy1.2 enrichment (Set 1): Cell pool was enriched for RGCs and macroglia using Thy1.2 and ACSA-2 (astrocyte cell surface antigen-2) antibodies respectively. Two retinas from 2 mice per age group were pooled for this method - 3-month (young), 12-month (middle-aged) and 24-month-old (old).         CD73 depletion (Set 2): The cell pool was depleted of rod photoreceptors using the CD73 antibody. Retinae from four 12-mo mice were prepared this way (2 retinas per pooled for each replicate).                                                                                     Thy1.2 enrichment and CD73 depletion (Set 3 and 4): Cell pool was split into two halves and each half was subjected to either Thy1.2 enrichment or CD73 depletion. Following this, the two portions were combined again. Two retinas from 2 mice per age group were used for this method - 3-mo (Set 3) and 12-mo (Set 4). Libraries were constructed according to manufacturer's protocol (10x Genomics Chromium Next GEM Single Cell 3' Kit v3.1 )