Anterior Eye Segment Culturing: A Method to Culture Cornea Bearing Anterior Segment of Porcine Eye
Published: April 30, 2023
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 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…
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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 | McMaster-Carr | 75165A675 |
