A novel filtration device for point of care preparation of cellular therapies

Abstract

Many biological samples are stored at cryogenic temperatures (i.e. temperatures below -1500C) to preserve their viability. Typically, these samples are stored in liquid nitrogen vapor-phase freezers. The underlying assumption is that biological samples show highly reduced degradation and metabolic activity while below Tg (the glass transition temperature). On the other hand, every time a sample is manipulated or temporarily removed from an LN2 freezer, the sample will experience thermal excursions marked by warm-up rates of several degrees per second. In these cases, the risk of harming a sample by inadvertently crossing the Tg threshold is very likely. This paper’s objective is to provide supporting data to fully characterize the thermal excursions of cryogenically frozen single vials filled with H2O during typical transient temperature events. Specifically, we focus our attention on the cold-chain steps that involve transferring a single vial from an LN2 vapor environment to either an ambient temperature or transportable dry ice (-800C) environment. In general, the warm-up rates experienced by the biological sample correlate to the thermal energy exchanged with the warmer environment via convective and conductive heat transfer. The magnitude of the heat transfer is driven by multiple factors: the warmer environment temperature and the overall time of exposure, the size and shape of the vial, its placement in a cryobox, the type of handling (manual or automated), the handling speed, etc. In this paper, we rationalize all the above factors and present experimental measurements supported by calibrated finite elements simulations showing typical expected warm-up rates. As a result, best-practice time constants for handling vials of biological samples without risking excessive thermal excursions above Tg are suggested.