Chicken Chorioallantoic Membrane-based Cancer Modeling: An In Ovo Model to Study Cancer Cell Tumorigenesis and Metastasis

Abstract

Source: Sharrow, A. C. et al. Using the Chicken Chorioallantoic Membrane In Vivo Model to Study Gynecological and Urological Cancers. J. Vis. Exp. (2020)

In this video, we describe the implantation of cancer cells onto a chicken chorioallantoic membrane to generate an in ovo model. The model successfully engrafts the ovarian cancer cells to study the progression of gynecological cancer.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Preparing the cancer cell suspension for transplantation (option 1)

NOTE: This is to be completed just prior to the implantation, which should ideally take place between days 7 and 10. Please see notes at the beginning of step 3 or 4 for further information concerning the implantation date. This approach was used for all the cell lines and cultured kidney cancer tumor digests.

1. Thaw the extracellular matrix solution on ice.....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
010 Teflon (PTFE) White 55 Duro Shore D O-Rings	The O-Ring Store	TEF010	Nonstick ring for cell seeding. 1/4"ID X 3/8"OD X 1/16"CS Polytetrafluoroethylene (PTFE).
Delicate Operating Scissors; Curved; Sharp-Sharp; 30mm Blade Length; 4-3/4 in. Overall Length	Roboz Surgical	RS6703	This is provided as an example. Any similar curved scissors would work as well.
Dremel 8050-N/18 Micro 8V Max Tool Kit	Dremel	8050-N/18	This kit contains all necessary tools
Fertilized chicken eggs (Rhode Island Red - Brown, Lab Grade)	AA Lab Eggs Inc.	N/A	A local egg supplier would need to be identified, as this supplier only delivers regionally.
Hovabator Genesis 1588 Deluxe Egg Incubator Combo Kit	Incubator Warehouse	HB1588D-NONE-1102-1588-1357	Other egg incubators may be used, but their reliability would need to be verified. After implantation, a cell incubator with the CO2 disabled may also be used.
Incu-Bright Cool Light Egg Candler	Incubator Warehouse	1102	Other candlers may be used; however, this is preferred among those that we have tested. This candler is included in the aforementioned incubator kit.
Matrigel Membrane Matrix HC; LDEV-Free	Corning	354248	Extracellular matrix solution
SKOV3	ATCC	HTB-77	Human ovarian cancer cell line.
Stirring Rods with Rubber Policeman; 5mm diameter, 6 in. length	United Scientific Supplies	GRPL06	This is provided as an example. Any similar glass stir rods would work as well.
Specimen forceps	Electron Microscopy Sciences	72914	This is provided as an example. The forceps used for pulling away the shell for bioluminescence imaging are approximately 12.8 cm long with 3 mm-wide tips.