title: GSM5047190: PBS; Rattus norvegicus; RNA-Seq
library_construction_protocol: Animals (5 animals for each condition) were anesthetized deeply with chloral hydrate, and a 5-mm section of the spinal cord centered at the injury site was dissected. Then, the tissue was chopped and incubated for 30 min at 37°C with 2 ml of collagenase (1 mg/ml in PBS) and centrifuged at 300 g for 5 min at 4°C. Two milliliters of 0.25% trypsin-EDTA solution was added to pelleted cells after collagenase removal and incubated at 37°C for 5 min. FBS (10%) was added to stop the digestion by trypsin, and the supernatant was removed by centrifugation. Precipitate dispersed in PBS with 0.5% BSA was strained through a 30-µm cell strainer twice. Cells were collected by centrifugation, and 10 ml of erythrocyte lysate was added. After 10 min, the cell suspension was centrifuged, and the cells were resuspended in PBS for counting. The single-cell suspension was then examined using the 10X Genomics Chromium Single Cell 3′ Reagent Kit v3 (10X Genomics) according to the manufacturer's protocol.