Renal Organotypic Culture on Chicken CAM: A Technique for Transplantation of Murine Kidney Explants on Chicken CAM to Induce Formation of Renal Vascular Architecture
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 Use transwell cell culture inserts designed for 6-well or 12-well plates. NOTE: Depending on the particular experiment, large or small minirese…
Divulgaciones
The authors have nothing to disclose.
Materials
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
Renal Organotypic Culture on Chicken CAM: A Technique for Transplantation of Murine Kidney Explants on Chicken CAM to Induce Formation of Renal Vascular Architecture. J. Vis. Exp. (Pending Publication), e20578, doi: (2023).