GSM7845000: AF-TD-FAW-1; Homo sapiens; RNA-Seq
Post-surgery, PDR fibrovascular membranes were placed in balanced salt solution (BSS) and stored on ice for transportation. For dissociation, membranes were transferred to a 500µL solution of 0.22µm filter sterilized 1x Dulbecco's Phosphate Buffered Saline (DPBS) with calcium and magnesium (Life Technologies, Carlsbad, CA) containing 2mg/mL Collagenase Type I (Worthington, Lakewood, NY). Samples were incubated at 370C for 30 minutes with gently trituration every 10 minutes. Samples were then spun down at 400g for 8 minutes at 40C and cell pellets were resuspended in 500µL 1X DPBS without calcium and magnesium (Life Technologies, Carlsbad, CA), 1% bovine serum albumen (BSA, citation), and 2mM EDTA and stored on ice until further processing. Single cell libraries were generated using a Chromium Single Cell 3' v3 kit (10x Genomics). RNA Quality and quantity were assessed using an Agilent Bio-analyzer. RNA-Seq libraries were prepared using Illumina mRNA TruSEQ kits following manufacturer's instructions.