GSM5939842: scRNA-seq corneal epithelium of diabetic monkey; Macaca fascicularis; RNA-Seq
We divided the cornea from anesthetized monkeys and the whole corneal was digested in the solution with Dispase II (10 mg/mL) at 37℃ for 2 hours. Next, the corneal epithelium was carefully dissected using forceps, and was transferred to 1.5 ml EP tube which contained the solution with Dispase II (2.5 mg/mL) at 37℃ for 15 minutes to dissociate the cell aggregates into single cells. EP tube was shaken gently every 5 minutes. Single cell suspensions were diluted at a concentration of about 1000 cells/μL in 0.04% BSA/Ames. Following the manufacturer's protocol, Single-cell 3' Library and Gel Bead Kit V2 (10x Genomics) and Gel Bead Kit (10x Genomics) were used for single-cell RNA-seq libraries preparation. Single cells were lysed and the released RNA was performed barcoded reverse transcription in GEMs. Sequencing libraries were sequenced on Illumina HiSeq X Ten.