Anterior Eye Segment Culturing: A Method to Culture Cornea Bearing Anterior Segment of Porcine Eye
Transcripción
For anterior eye segment culturing, begin with a suitable culture dish. The culture dish has an elevated stage at the center, connected to two fluid ports for inflow and outflow.
First, place a fresh porcine anterior eye segment containing cornea over the stage. Subsequently, place a clamp ring over the eye segment and secure it. Next, prepare a perfusion set-up comprising a syringe filled with a suitable culture medium. Connect the syringe to a valve, filter, and tubing with a bent needle hub at the other end.
Prepare a similarly assembled reservoir set-up using an empty syringe. Now, connect the needle hubs of the perfusion and the reservoir set-up to the inflow and outflow ports, respectively. Gently perfuse the media and allow the anterior segment to inflate.
Continue the perfusion and allow the bubbles and excess medium to flow into the reservoir. Thereafter, incubate the culture dish at the desired height and standard physiological conditions. This results in a three-dimensional model that simulates the complex architecture of the eye under physiological conditions.
Connect a pressure transducer to the reservoir valve to constantly monitor the eye pressure. The anterior eye segment is now ready to use for further experiments.
