Nod scid

For assessment of diabetes, female NOD/ShiLtJ mice obtained from The Jackson Laboratory (Bar Harbor, ME) were used. For evaluation of insulitis and for adoptive transfer study, NOD/ShiJcl and NOD-SCID mice obtained from CLEA Japan Inc. (Tokyo, Japan) were used. All the mice were maintained under specific pathogen-free conditions, housed in autoclaved cages, and provided with autoclaved food and water. The studies conducted at The Jackson Laboratory were performed according to an IACUC-approved protocol and in compliance with the Guide for the Care and Use of Laboratory Animals. The experiments conducted at Tokyo Women's Medical University were performed according to a protocol approved by internal and external committees and were in compliance with the prescribed guidelines.

Animal experiments were performed in accordance with a protocol approved by the Animal Care and Use Committee of the Faculty of Medicine, Tokyo Medical University.
Eight-week-old NOD.CB17-Prkdc^scid/J (NOD SCID) male mice (CLEA Japan, Tokyo, Japan) were employed for the fracture model. Nine mice were used for each group. Each mouse was anesthetized with medetomidine (0.75 mg/kg), midazolam (4.0 mg/kg), and butorphanol tartrate (5.0 mg/mL) via intraperitoneal injection. A skin incision was made in the left tibia and a blunt dissection of the muscle was made to expose the tibia. Osteotomy was performed using a diamond cutting disc at the mid-diaphysis. The bone marrow cavity of the fractured tibia was stabilized by inserting a 23 G spinal needle (TOP, Tokyo, Japan) [36 (link),37 (link)]. DPSC sheets were cultured on temperature-responsive dishes in NM or OM with or without TH for 14 days. Then, the DPSCs were stained with PKH26 (Sigma-Aldrich, Darmstadt, Germany) in accordance with the manufacturer’s protocol. DPSC sheets were removed at room temperature for 30 min, and then transplanted so that they were wrapped around the tibia like a bandage on the fracture line. The incision was closed with 6-0 nylon sutures, and radiography was conducted to confirm the fracture site. After mice were euthanized on day 14, the fractured tibiae were harvested.