GSM8187376: PFC, BCAS, Female, 3; Mus musculus; RNA-Seq
Mice were anesthetized with isoflurane and transcardially perfused with ice-cold, bubbled artificial cerebrospinal fluid (ACSF). The brain was dissected and mounted for sectioning. Coronal brain sections were generated with a Compresstome vibrating microtome. The vibratome chamber was filled with a sucrose-based cutting solution, and sections were cut to a thickness of 250 µm. Sections were subsequently transferred into ice-cold ACSF-Trehalose (ACSFT). Prefrontal cortex regions were excised from individual brain. Tissue were digested with Pronase (Sigma-Aldrich, 537088; 2 mg/ml) diluted in bubbled ACSFT at room temperature for 45 minutes. Subsequently, tissue was washed with 1% FBS in ACSFT three times. Mechanical dissociation of tissue into a single-cell suspension was achieved by pipetting the samples using pipette tips of decreasing bore sizes (p1000, p200, p100, p20). The resultant cell suspensions were filtered through a 0.2 µm cell strainer. 10X Chromium Next GEM Automated Single Cell 3ʹ (v3.1 Chemistry).