In this video, we demonstrate the culturing method of a cornea-bearing anterior eye segment from a porcine eye. This culture system serves as an experimental model to study eye injuries.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Setting up anterior segments in organ culture dishes Place a single anterior segment (AS) in a Petri dish with AS inverted (cup up). Using a cotton swab, wet in the media and gently dab in the center of the cornea to remove any pigment. Using forceps to hold the eye and the same swab, wipe the cotton swab around the sclera to remove ext…
Divulgations
The authors have nothing to disclose.
Materials
10-32 Polycarbonate straight plug, male threaded pipe connector
McMaster-Carr
51525K431
10-32 Socket cap screw, ½"
McMaster-Carr
92196A269
10 mL syringe
BD
302995
20 mL syringe
BD
302830
Anti-Anti
Gibco
15240-096
Ball-End L key
McMaster-Carr
5020A25
BD Intramedic PE 160 Tubing
Fisher Scientific
14-170-12E
Cotton swabs
Puritan
25-8061WC
DMEM media
ATCC
30-2002
FBS
ATCC
30-2020
Fine forceps
World Precision Instruments
15914
Glutamax
Gibco
35050-061
High temperature silicone O-ring, 2 mm wide, 4 mm ID
McMaster-Carr
5233T47
Nail (puncture object)
McMaster-Carr
97808A503
Nylon syringe filters
Fisher
09-719C
PBS
Gibco
10010-023
Polycarbonate, three-way, stopcock with male luer lock
Fisher
NC9593742
Stainless steel 18 G 90 degree angle dispensing needle
McMaster-Carr
75165A81
Stainless steel 18 G straight ½'' dispensing needle