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摘要
To study mucosal immunity and conduct HIV vaccine trials, it is important to be able to cryopreserve mucosal specimens and recover them in functional viable form. Obtaining a good recovery depends, in part, on cooling the cells at the appropriate rate, which is determined by the rate of water transport across the cell membrane during the cooling process. In this study, the cell membrane permeabilities to water at subzero temperatures of human vaginal mucosal T cells and macrophages were measured using the differential scanning calorimetry method proposed by Devireddy et al. in 1998. Thermal histograms were measured before and after cell lysis using a Slow-Fast-Fast-Slow cooling program. The difference between the thermal histograms of the live intact cells and the dead lysed cells was used to calculate the temperature-dependent cell membrane permeability at subzero temperatures, which was assumed to follow the Arrhenius relationship, , where L-pg is the permeability to water at the reference temperature (273.15 K). The results showed that L-pg=0.0209 +/- 0.0108m/atm/min and E-a=41.5 +/- 11.4kcal/mol for T cells and L-pg=0.0198 +/- 0.0102m/atm/min and E-a=38.2 +/- 10.4kcal/mol for macrophages, respectively, in the range 0 degrees C to -40 degrees C (mean +/- standard deviation). Theoretical simulations predicted that the optimal cooling rate for both T cells and macrophages was about -3 degrees C/min, which was proven by preliminary immune cell cryopreservation experiments.
- 出版日期2016-8
- 单位中国科学技术大学
