GSM6384092: Non-injured/MTZ; Danio rerio; RNA-Seq
The telencephalon was dissected in ice-cold Hank's Balanced Salt Solution (HBSS). Seven telencephalons per each experimental condition were pooled and processed. Isolated telencephalons were chemically dissociated for 30 - 40 min at room temperature in HBSS containing papain (20U/ml) and DNase (2000U/ml) with gentle trituration every 10 minutes. The cell suspension was filtered with a 40 μm cell strainer, then 10% FBS in L-15 medium was added to stop the enzymatic reaction. The dissociated cells were washed with 10% FBS/PBS solution, and the final pellet was resuspended in 10% FBS/PBS for cell sorting. Before cell sorting, cells were again passed through a 40μm cell strainer, and Sytox blue dead cell stain was added to label the dead cells. FACS sorting was performed using BD FACSAria II, and  9000 to 20000 dTomato+ cells were collected in a 10% FBS/PBS precoated tube. Single-cell RNAseq libraries were prepared using Chromium Single Cell 3' Reagents Kits v3 according to the manufacter's instructions with 12 cycles of cDNA amplification and 15 cycles of indexing PCR