Isothermal Vitrification Methodology Development for Non-Cryogenic Storage of Archival Human Sera

摘要: Biorepositories worldwide collect human serum samples and store them for future research. Currently, hundreds of biorepositories across the world in refrigerators, freezers, or liquid nitrogen without following any specific cryopreservation protocol. This method storage is both expensive potentially detrimental to biospecimens. To decrease cost freeze/thaw stresses, we explored feasibility storing archival at non-cryogenic temperatures using isothermal vitrification. When biospecimens are vitrified, biochemical reactions can be stopped, specimen ceases degrade, macromolecules stabilized requiring cryogenic storage. In this study, 0.2, 0.4, 0.8 M trehalose; 0, 0.005 0.01 dextran; 0 10% (v/v) glycerol was added samples. The were either dried diffusively as sessile droplets desiccated under vacuum after they adsorbed onto glass microfiber filters. transition (Tg) measured by temperature-ramp Fourier Transform Infrared (FTIR) spectroscopy. Sera vitrified 4 ± 2 °C when trehalose dextran two hours. Western immunoblotting showed that proteins minimally degraded stored up one month °C. About 80% all recovered on filters, recovery did not with time. These results demonstrated long-term hypothermic (and non-cryogenic) temperatures.