The mice were injured as previously described [39 (link),40 (link),41 (link),42 (link),43 (link)]. Briefly, ketamine (100 mg/kg), xylazine (10 mg/kg), and buprenorphine SR (0.5 mg/kg) were administered subcutaneously for anesthesia and analgesia before surgery. Hair on the scalp was removed. The mice were then positioned in a stereotaxic frame at 37 °C using a homeothermic blanket system (Harvard apparatus, Lewes, DE, USA). A Φ = 4 mm craniectomy was drilled over the right parietal bone (−2.5 mm A/P and 2.0 mm lateral from bregma), and injury was induced at the center of the craniectomy using a Φ = 3 mm flat tip connected to an eCCI-6.3 device (Custom Design & Fabrication, LLC, Petersburg, VA, USA) at a velocity of 5.0 m/s, with a 250 ms impact duration and a depth of 2.0 mm (n = 20) or 2.5 mm (n = 23, 15 of which were used for astrocyte isolation). Two severely injured mice (2.5 mm depth) died unexpectedly during the course of the study and were therefore excluded. Kwik-Sil (WPI, Sarasota, FL, USA) was applied to cover the craniectomy, and the incision was closed with 4.0 PDO sutures (AD surgical, Sunnyvale, CA, USA).
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