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  • Protocol to quantify enzymatic effects on vitreous liquefaction in porcine eyes using a transwell-plate system.

Protocol to quantify enzymatic effects on vitreous liquefaction in porcine eyes using a transwell-plate system.

STAR protocols (2022-10-09)
Julian Wolf, Luís E Sabage, Young Joo Sun, Vinit B Mahajan
ABSTRACT

This protocol describes an ex vivo model to quantify enzymatic effects on vitreous liquefaction using porcine eyes in a transwell-plate system via induced syneresis. It provides a standardized dissection process and performs critical steps for gel-liquid separation with high precision, minimal tissue loss, and scalability. The protocol can be applied to other studies investigating vitreous liquefaction or gelatinous tissue analysis and can also serve to study vitreous liquefaction in vivo as it may occur during aging or disease progression.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Collagenase from Clostridium histolyticum, 0.2 μm filtered, suitable for release of physiologically active rat epididymal adipocytes, Type II-S, 0.5-5.0 FALGPA units/mg solid, ≥125 CDU/mg solid
Sigma-Aldrich
Trypsin from porcine pancreas, lyophilized powder, Type II-S, 1,000-2,000 units/mg dry solid
Sigma-Aldrich
Hyaluronidase from bovine testes, Type I-S, lyophilized powder, 400-1000 units/mg solid