Chick Chorioallantoic Membrane or CAM Tumor Model: An In Ovo Method to Simulate Perineural Invasion of Tumor Cells
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 describe the method to co-culture dorsal root ganglia and cancer cells on chick chorioallantoic membrane or CAM to study perineural invasion.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Grafting DRG on the CAM (estimated timing: 40 min, day 8) Prepare a 6 cm culture dish with HBSS medium to wash the DRGs before implantation. Bring the prepared eggs to the cell culture laminar flow cabinet. Remove the paraffin membrane from the egg (Figure 1A). With fine sterile forceps, …
Representative Results
Figure 1: Grafting of DRG, cells, and harvesting of CAM: On day 8: A. CAM easily observed after paraffin wax membrane removal. B-C. With fine forceps, DRG is placed onto the CAM. D. Egg is covered with film dressing and put in the incubator; arrows point to the openings that are covered. On day 10: E-F….
Divulgaciones
The authors have nothing to disclose.
Materials
| Egg incubator | GQF Digital Sportsman | #1502 | Egg incubator equipped with automatic rotator digital thermostat temperature and humidity controls |
| Engraving cutter | DREMEL | #108 | Used to drill the egg shell |
| Extra fine Graefe forceps, curved | Fine Science Tools (FST) | #11151-10 | Used to graft DRG on to the CAM on day 8 and to harvest CAMtissue on day 17 |
| Extra fine Graefe forceps, straight | Fine Science Tools (FST) | #11150-10 | Used to graft DRG on to the CAM on day 8 and to harvest CAM tissue on day 17 |
| Fertilized Lohmann White Leghorn eggs | Fertilized eggs at early fertilization days preferably on first day postfertilization. Eggs used in this protocol are from Michigan State University Poultry Farm. | ||
| Filter Forceps | EMD Millipore | #XX6200006P | Blunt forceps used to remove the egg shell |
| HBSS (1x) | Gibco | #14025-092 | Hank`s Balanced Salt Solution |
| Paraffin wax membrane | Parafilm laboratory film | #PM-996 | Used to temporarily cover the egg openings until DRG grafting on day8 |
| PBS (1x) pH 7.4 | Gibco | #10010-023 | Phosphate Buffered Saline |
| Tegaderm Transparent Film Dressing | 3M | #9505W | Sterile, 6x7cm used to cover the egg openings during incubation |
Tags
Citar este artículo
Chick Chorioallantoic Membrane or CAM Tumor Model: An In Ovo Method to Simulate Perineural Invasion of Tumor Cells. J. Vis. Exp. (Pending Publication), e20477, doi: (2023).