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2.3 Preparation of sperm suspensions On February 24 th , 2024, frogs were euthanised according to methods approved by the University of Wollongong s Animal Ethics Committee (see section 2.8 Ethics review statement below), and testis tissue immediately extracted, weighed, and macerated in 170L of simplified amphibian ringer (SAR: 113mM NaCl, 1mM CaCl 2 , 2mM KCl, 3.6mM NaHCO 3 )

Plasma concentrations of GHK decline with age, from roughly 200 ng/mL in young adults to about 80 ng/mL by age 60 (Pickart, Vasquez-Soltero & Margolina, BioMed Research International, 2015)