Sperm cryopreservation is a valuable tool to support preservation and management of valuable genetic resources. Cryopreservation of sperm requires the balance of multiple steps such as the cooling rate and method. The cooling step is commonly performed using a programmable freezer that provides a precisely controlled temperature profiles for cryopreservation. However, controlled-rate freezers can cost $15,000-$50,000, and most breeding and research programs cannot afford this equipment. Three-dimensional (3-D) printed open-hardware devices offer a low-cost alternative to programmable freezers while providing easy fabrication and standardizable reproducibility. In this study, cooling rates and post-thaw sperm quality processed by two open-hardware devices developed at the AGGRC were compared with a commercial programmable freezer (IceCube, Minitube). Both devices, designed for 0.25-ml and 0.5-ml French straws, provided various predetermined cooling rates. The Cajun Ejector (Figure 1A) utilized vertical freezing inside a standard nitrogen vapor shipping dewar. Different cooling rates can be set by adjusting the height of the straw holder into the shipping dewar. The CryoKit (Figure 1B), a horizontal rack was designed to float on liquid nitrogen inside of a Styrofoam box, was set for different cooling rates by adjusting the height of samples above liquid nitrogen, and with different 3-D printed configurations. Testes from five male Xenopus laevis were collected and fresh quality was assessed by estimating sperm concentration, percentage of viable sperm and percentage of motile sperm. Sperm concentration was adjusted to 1.6 ×108 cells/mL, samples were diluted 1:1 with a cryoprotectant (10% dimethylformamide and 10% sucrose), and 100µL loaded into straws and cooled at -10C°/min using the three different devices. Then, samples were thawed by immersion in a water bath at 40ºC for 5s and sperm quality was assessed and compared among the devices. Preliminary results (n=2) showed a similar cooling rate between the programmable freezer and the Cryokit (-10±1ºC/min) and a faster rate when using the Cajun Ejector(>-40°C/min). Consequently, there were no differences between the sperm viability using the programmable freezer (35±2.5%) and the Cryokit (45±7.1%) but lower sperm survival was obtained when using the Cajun Ejector (17±4.2%). Communities performing cryopreservation of amphibian sperm with biomedical models such as Xenopus or endangered species could benefit from use of these portable, inexpensive, and reproducible devices.