title: Determination of the Membrane Permeability to Water of Human Vaginal Mucosal Immune Cells at Subzero Temperatures Using Differential Scanning Calorimetry.
abstract: 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, [Formula: see text], where Lpg is the permeability to water at the reference temperature (273.15 K). The results showed that Lpg = 0.0209 ± 0.0108 μm/atm/min and Ea = 41.5 ± 11.4 kcal/mol for T cells and Lpg = 0.0198 ± 0.0102 μm/atm/min and Ea = 38.2 ± 10.4 kcal/mol for macrophages, respectively, in the range 0°C to -40°C (mean ± standard deviation). Theoretical simulations predicted that the optimal cooling rate for both T cells and macrophages was about -3°C/min, which was proven by preliminary immune cell cryopreservation experiments.
authors.affiliation: 1 Department of Mechanical Engineering, University of Washington , Seattle, Washington.|2 School of Mechanical and Materials Engineering, Washington State University , Everett, Washington.
authors.forename: Zhiquan
authors.initials: Z
authors.lastname: Shu
authors.affiliation: 3 Department of Obstetrics and Gynecology, University of Washington , Seattle, Washington.
authors.forename: Sean M
authors.initials: SM
authors.lastname: Hughes
authors.affiliation: 1 Department of Mechanical Engineering, University of Washington , Seattle, Washington.
authors.forename: Cifeng
authors.initials: C
authors.lastname: Fang
authors.affiliation: 1 Department of Mechanical Engineering, University of Washington , Seattle, Washington.
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Hou
authors.affiliation: 4 Department of Electronic Science and Technology, University of Science and Technology of China , Hefei, China .
authors.forename: Gang
authors.initials: G
authors.lastname: Zhao
authors.affiliation: 3 Department of Obstetrics and Gynecology, University of Washington , Seattle, Washington.
authors.forename: Michael
authors.initials: M
authors.lastname: Fialkow
authors.affiliation: 3 Department of Obstetrics and Gynecology, University of Washington , Seattle, Washington.
authors.forename: Gretchen
authors.initials: G
authors.lastname: Lentz
authors.affiliation: 3 Department of Obstetrics and Gynecology, University of Washington , Seattle, Washington.|5 Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center , Seattle, Washington.
authors.forename: Florian
authors.initials: F
authors.lastname: Hladik
authors.affiliation: 1 Department of Mechanical Engineering, University of Washington , Seattle, Washington.
authors.forename: Dayong
authors.initials: D
authors.lastname: Gao
mesh_terms: D001692:Biological Transport; D002152:Calorimetry, Differential Scanning; D002463:Cell Membrane Permeability; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D006801:Humans; D018928:Immunity, Mucosal; D008264:Macrophages / Q000166:cytology*; D009092:Mucous Membrane / Q000166:cytology / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology*; D046509:Tissue Culture Techniques; D014621:Vagina / Q000166:cytology*; D014867:Water / Q000378:metabolism*
chemical_list: D014867:Water