GSM5770419: mouse RPE-choroid complex AAV-GFP rep1; Mus musculus; RNA-Seq
To isolate viable single cells from RPE/choroid tissue, the mice were euthanized and then enucleated one eye for further processing. Each eye was cleaned from extraocular tissue and a circumpherencial incision was performed around the limbus cornea to remove the anterior segment, including cornea, lens, iris and ciliary body. The retina was carefully separated from the choroid, then each RPE-choroid tissue was cut into eight petals.  RPE/choroid tissue was digested with 200μL of solution containing 52U/ml Papain, Suspension (Worthington,LS003126),3mg/ml DNase I (BioFroxx,1121MG010) at 37°C for 15 min. Single cell suspension was created by pipetting up and down for several times with 10μL pipette , then filtered through a 45-µm cell strainer before being loaded onto the 10x Genomics Chromium instrument. The scRNA-seq libraries were generated using the 10x Genomics Chromium Single Cell v3 reagent kit according to the manufacturer's instructions