Renal Organotypic Culture on Chicken CAM: A Technique for Transplantation of Murine Kidney Explants on Chicken CAM to Induce Formation of Renal Vascular Architecture

Abstract

Source: Kaisto, S. et al. Optimization of Renal Organoid and Organotypic Culture for Vascularization, Extended Development, and Improved Microscopy Imaging. J. Vis. Exp. (2020)

This video describes a method for studying organ development using an improved xenotransplantation setup on chorioallantoic membrane (CAM) from avian embryos. This method allows for the development of fully vascularized cultured embryonic organs and organoids.

Protocol

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

1. Fabrication of Minireservoirs for Cultivation of Mouse Embryonic Kidneys and Renal Organoids on Chicken CAM and Setting Up Xenotransplantation Experiments

1. Use transwell cell culture inserts designed for 6-well or 12-well plates.
   NOTE: Depending on the particular experiment, large or small minireservoirs can be used. It is best to use small minireservoirs for xenografting up to eight embryonic kidneys or when several minireservoirs will be place....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Cell Incubator	Panasonic	13090543	Temperature set at 37° C, 90% humidity, 5% CO2
Cell Incubator	SANYO	10070347	Chicken CAM-culture incubator. Temperature set at 37° C, 90% humidity, 5% CO2
Dulbecco's Modified Eagle's Medium	Sigma	D777	High glucose
Ethanol (70%)	VWR
Fetal Bovine Serum	HyClone	SH3007003HI Thermo Scientific
Forceps DUMONT #5	Dumont	#5SF
PBS -/-	Corning	20-031-CV
PBS +/+	Biowest	X0520-500	Washing the mini reservoirs.
Penicillin and Streptomycin	Sigma	P4333
Thincert 12-well Cell Culture Inserts with 0.4 µm-pore Polystyrene Membrane	Greiner Bio-One	665641
Thincert 6-well Cell Culture Inserts with 0.4 µm-pore Polystyrene Membrane	Greiner Bio-One	657610