DS allows the morphology of the ice interface to be varied under conditions where the local chemical conditions of the residual solution can be kept constant, which is different to what happens in PS where progressive exclusion of both solutes and, in some situations, cells occurs ahead of the ice front [11]. DS also allows better homogeneity of the cooling profile throughout the entire sample, whereas, as seen here, PS results in differential thermal profiles towards the sample centre as the excluded solutes, generating areas of local undercooling, result in variable release of latent heat of ice crystal formation which have to be dissipated from the sample Selleckchem Daporinad core before controlled cooling can proceed.

However, for large cell masses contained within an irregular geometry as investigated here, engineering a DS approach to cryo-cooling would prove to be challenging. In the current work, solidification proceeded only through static surface cooling conditions, with ice growth primarily determined by the thermal properties and 3-dimensional structure of the sample. Another high throughput screening compounds factor worthy of comment is that the experimental systems used here had little excess cryoprotectant additive and there would be little settling effect of ELS on the ice crystal progression

– all the samples were in effect ‘settled’ by removing the extra CPA volume. The process of ice propagation in this system may differ compared with conventional cell and protein suspensions, where sedimentation of cells may occur before initiation of freezing and, secondly, cells and proteins may be pushed ahead of ice fronts during progressive solidification. While success has been reported with large volumes in flat bag cryopreservation, these have generally been deliberately

compressed into a thin wafer or ‘slab’ format with little internal temperature gradients and so often experience NS. It is possible to observe PS in bags however, if the bag temperature is not thermally equilibrated prior to the onset of solidification [15] and [25]. Such flat-bag approaches would be very difficult to adapt for BAL cryopreservation due to the geometries Verteporfin involved, where the end-product would ideally reside in a cylindrical fluidised bed format. The varying temperature profiles throughout the sample when cooling a large cylinder have been recognized for some time [19]. Previous studies have shown that the level of freeze-concentration of solutes is dependent on the cooling rate and this has been studied in detail in cylindrical vessels [13]. In cylindrical configurations, the solutes increased in concentration radially from the edge of the cylinder to the centre, and this was accompanied by aggregation of some proteins within the core layers. Due to the alginate sphere composition of the test BAL, cell aggregation will not occur here as the cells are already immobilised.