GSM8757797: Smoke; Mus musculus; RNA-Seq
Choroid-sclera and retina were digested separately in a papain solution (40U papain / 10mL DPBS) for 45 minutes. Papain was then neutralized in a trypsin inhibitor solution (0.15% ovomucoid in DPBS) and the tissue was triturated to generate a single-cell suspension. Following trituration, the cells were pelleted, resuspended, and filtered through a 40μm Nitex mesh filter to eliminate any clumped cells. Dissociated cells from retinas were finally incubated with PE anti-mouse CD73 (127206, Biolegend, US) followed by anti-PE microbeads (130-048-801, Miltenyi Biotec, US) to deplete rods. Single cell suspensions from retina and choroid-sclera were mixed and diluted at a concentration of 200 cells/μL in 0.2% BSA/DPBS for loading into 10x Genomics Chromium. Droplet generation and cDNA libraries were completed by Novogene company (Beijing,China).