title: GSM9010623: Ctrl_14d; Mus musculus; RNA-Seq
library_construction_protocol: For calvarial defect creation, anesthesia was performed with 2-3% isoflurane in 100% oxygen at a flow rate of 1 L/min and animals were operated upon on a warm animal surgery station. Firstly, hair overlying the calvaria was clipped, and a 4 mm skin incision was made over the right frontal bone. Next, a 1.8 mm diameter full thickness circular defect was created in the non-suture associated frontal bone using a micro surgical drill and a trephine drill bit (Xemax Surgical, Napa Valley, CA). Meticulous care was taken not to injure the underlying dura mater. Finally, the skin was sutured and the animal was monitored per established postoperative protocols. Post-operative monitoring was performed in accordance with institutional policy. Skulls were harvested up to 7d and 14 d after defect, respectively. Library preparation and sequencing were conducted by Majorbio Bio-Pharm Technology Co. Single-cell suspensions were loaded onto the Chromium Single Cell Controller (10x Genomics) to generate single-cell gel bead emulsions, following the manufacturer's guidelines (Chromium Single Cell 3ʹ v3.1). The Chromium Single-Cell 3' Gel Bead and Library V3 Kit was used for cDNA synthesis and amplification, and sequencing was carried out on an Illumina NovaSeq 6000 platform.