Harvesting DRG from Rat Spine: A Procedure to Extract Dorsal Root Ganglion from the Spine of a Rat Model
Published: April 30, 2023
Abstract
Source: Schmidt, L. B. et al. The Chick Chorioallantoic Membrane In Vivo Model to Assess Perineural Invasion in Head and Neck Cancer. J. Vis. Exp. (2019)
In this video, we demonstrate the extraction of dorsal root ganglion or DRG from a harvested rat spine for downstream applications.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Harvest and Preparation of DRGs (estimated timing: 2 h, day 8) NOTE: Experiments with mice and rats require approval from the IACUC. In some countries, the use of chicken eggs also requires approval. Get six to seven week-old Sprague Dawley rats (~200 g in weight) to extract …
Representative Results
Figure 1: DRG extraction on day 8. A. Rat schematic illustrating the anatomical location of the spine. B. Diagram of the rat vertebrae configuration showing different body regions; green for cervical, dark blue for thoracic, orange for lumbar and light blue for sacral vertebrae. C-D. Ventral aspect of the rat spine after surgical exci…
Offenlegungen
The authors have nothing to disclose.
Materials
| PBS (1x) pH 7.4 | Gibco | # 10010-023 | Phosphate Buffered Saline |
| Sprague Dawley rats (females) | Charles River laboratories | Strain code: 001 | 6-7 weeks old (190-210g in weight) |
| Dumont # 5 fine forceps | Fine Science Tools (FST) | # 11254-20 | Used to harvest DRG |
| Extra fine Graefe forceps, curved | Fine Science Tools (FST) | # 11151-10 | Used to graft DRG onto the CAM on day 8 and to harvest CAM tissue on day 17 |
| Extra fine Graefe forceps, straight | Fine Science Tools (FST) | # 11150-10 | Used to graft DRG onto the CAM on day 8 and to harvest CAM tissue on day 17 |
| DMEM (1x) | Gibco | # 11965-092 | Dulbeecco`s Modified Eagle Medium |
| CellTracker Red CMTPX fluorescent dye | Life Technologies | # C34552 | Reconstitute 50µg in 40µL of DMSO and stock at -20oC. Use 1µL of stock solution/mL of culture medium |
| Pen/Strep | Gibco | # 15140-122 | 10,000 Units/mL Penicilin, 10,000 µg/mL Streptomycin |
| HI FBS | Gibco | # 10082-147 | Heat-inactivated Fetal Bovine Serum |
| DMSO | Fisher Bioreagents | # BP231-100 | Dimethyl Sulfoxide |
Tags
Diesen Artikel zitieren
Harvesting DRG from Rat Spine: A Procedure to Extract Dorsal Root Ganglion from the Spine of a Rat Model. J. Vis. Exp. (Pending Publication), e20476, doi: (2023).