Overview
This video demonstrates a surgical procedure to dissect the palate tissue from a murine model.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Dissection of Palate Tissue from Adult Mouse
- Sacrifice an adult C57BL/6J mouse (either male or female) by cervical dislocation in compliance with the facility's regulations relating to animal welfare.
- Remove the hair around the mouth with a shaver. Using scissors, cut from the cheek toward the jaw, on both sides.
NOTE: The mouse needs to be anesthetized before sacrifice. An anesthetic mixture of medetomidine, midazolam, and butorphanol is used (see Table of Materials).
- Remove the hair around the mouth with a shaver. Using scissors, cut from the cheek toward the jaw, on both sides.
- Use forceps to open the mouth wide and absorb any blood using a cotton swab. To disinfect the palate, wipe the inside of the mouth with a cotton swab containing 10% povidone-iodine.
- To harvest the mouse palate, first, use a surgical scalpel blade to make a full-thickness marginal incision along the palate side of the maxillary teeth. Then, carefully dissect the entire palate using a raspatorium.
NOTE: A raspatorium is a tool used to elevate a mucoperiosteal flap (see Table of Materials) (Figure 1B). - Quickly transfer the palate tissue to a 15 mL tube containing 4 mL of complete medium + antibiotic-antimycotic solution. Keep the tissues on ice until ready for incubation.
NOTE: For collection from multiple mice, palate tissues may be kept on ice for up to 4 h.
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Representative Results
Figure 1: Overview of the dissection procedure and isolation of mouse oral keratinocytes. (A) Schematic representation of the mouse oral cavity. (B) Instruments used to dissect the palate and isolate mouse oral keratinocytes. (C) Brightfield image of the mouse palate. Scale bar: 100 µm. (D) Summary of the protocol.
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Materials
| Name | Company | Catalog Number | Comments |
| Iris Scissors | Muromachi Kikai | 14090-09 | |
| Medetomidine | Nippon Zenyaku Kogyo | Domitor | |
| Midazolam | Astellas Pharma | Dormicum | |
| No.15 Disposable scalpel | Feather | 219AABZX00136000 | |
| Butorphanol | Meiji Seika Pharma | Vetorphale | |
| Half-curved forceps | BRC Bio Research Center | PRI13-3376 | |
| Povidone-iodine | Y's Square | 872612 | |
| Raspatorium #14 | AS ONE | 8-4599-01 | |
| Fine forceps | BRC Bio Research Center | PRI13-3374 | |
| Antibiotic-Antimycotic 100x | Gibco | 15240062 | |
| 15 mL sterile conical tube | Falcon | 352096 | |
| Cotton | AS ONE | 63-1452-97 | |
| EpiLife Defined Growth Supplement (EDGS) | Gibco | S0120 |
