Anterior Eye Segment Culturing: A Method to Culture Cornea Bearing Anterior Segment of Porcine Eye

Abstract

Source: Boice, E. N., Snider, E. J. Anterior Segment Organ Culture Platform for Tracking Open Globe Injuries and Therapeutic Performance. J. Vis. Exp. (2021).

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

1. 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 extra pigment.
2. Invert the AS and place on top of bottom part of the dish over the elevated region, centering the cornea over the elevated regio....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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	McMaster-Carr	75165A675