C3fe swv mbpshi j

Shiverer mice (C3Fe.SWV-Mbpshi/J, The Jackson Laboratory, USA) lack myelin basic protein and myelin in the CNS^{39 (link),40 (link)}. Shiverer mice 8–10 weeks old (N = 3, each group) were anesthetized and injected with 50,000 iPSC derived Zfp488-ZsGreen1/PDGFR-α⁺ OPCs or sham injected with the same volume of phosphate buffered saline into the corpus callosum (surgical procedure and stereotaxic coordinates were identical with the LPC injections coordinates). Mice were administered cyclosporine 10 mg/kg, intraperitoneally daily for 8 weeks for immune suppression after which they were sacrificed. Brains were processed for electron microscope imaging. One-micron meter sections of the injected areas were analyzed for the formation of the multilayered compact myelin sheath. We could not derive enough control OPCs (with the empty virus) for transplantation using this protocol. All animal procedures were approved by the IACUC committee of UC Davis. All experiments (in vitro and invivo including supplementary information) were performed in accordance with relevant guidelines and regulations.

Shiverer mice (C3Fe.SWV-Mbpshi/J, The Jackson Laboratory, USA) lack MBP and compact myelin in the CNS. Therefore, these mice serve as a good model to test the functional capacity of transplanted OPCs toward forming MBP-positive compact myelin sheath around axons. Shiverer mice 8–10 weeks old (N = 3; 2 males, 1 female) were injected with 50,000 iPSC-derived GANT61-OLIG2⁺/GFP⁺/PDGFR-α⁺ OPCs. Mice were deeply anesthetized by a cocktail of ketamine (70 mg/kg) and xylazine (10 mg/kg). Then, 1 μL of cell suspension was injected aseptically (at the rate of 0.3 μL/min) into the corpus callosum using a Hamilton syringe at the following stereotaxic coordinates: anteroposterior 0.2, mediolateral 1.1 with Bregma as a reference, and dorsoventral 2 mm from the skull surface. Mice were administered cyclosporin 10 mg/kg daily intraperitoneally for immune suppression, beginning 2 days before cell transplantation until sacrifice. Postoperative care was given according to the Institutional Animal Care and Use Committee (IACUC) protocols approved by the University of California, Davis. Mice were sacrificed after 4 or 6 weeks, and the corpus callosum sections (2–3 mm) containing the injection site was microdissected and processed for imaging under an electron microscope or for confocal imaging.