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Merck

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

STAR protocols (2022-10-09)
Julian Wolf, Luรญs E Sabage, Young Joo Sun, Vinit B Mahajan
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This protocol describes an exย 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 inย vivo as it may occur during aging or disease progression.

MATERIALS
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Sigma-Aldrich
Collagenase from Clostridium histolyticum, 0.2 ฮผm filtered, suitable for release of physiologically active rat epididymal adipocytes, Type II-S, 0.5-5.0 FALGPA units/mg solid, โ‰ฅ125 CDU/mg solid
Sigma-Aldrich
Hyaluronidase from bovine testes, Type I-S, lyophilized powder, 400-1000 units/mg solid
Sigma-Aldrich
Trypsin from porcine pancreas, lyophilized powder, Type II-S, 1,000-2,000 units/mg dry solid