NSPC Grafting in Hippocampus

NSPC grafting was performed as described previously (18 (link)). Mice were mounted onto a stereotactic head holder (Kopf Instruments, Tujunga, CA, USA) under anesthesia with isoflurane (Isoba® vet; Schering-Plough Corp., NJ, USA; 5% for induction, and 2% to 3% for maintenance) in the flat skull position. In order to graft NSPCs into the hippocampus, a 5-μL 26 gauge syringe (Innovative Labor Systeme, Stuetzerbach, Germany) attached to the holder was inserted according to the following coordinates: body weight < 9 g: 0.45 × (distance from lambda to bregma) mm posterior and ± 1.2 mm lateral to bregma, 3.0 mm deep from the skull surface; body weight ≥ 9 g, 0.42 × (distance from lambda to bregma) mm posterior and ± 1.3 mm lateral to bregma, 3.2 mm deep from the skull surface. Before the needle was inserted, a small hole was drilled in the proper position according to the above coordinates. Then, 1 × 10⁵ NSPCs in 2 μL DMEM were injected very slowly over a 2-min period, with a 4-min delay prior to removal of the syringe, and 2 min were allowed for syringe removal. No immunosuppressive drugs were administered.