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Initial measurement of the intracellular glass transition temperature of a mammalian cell type under cryopreservation conditions

Initial measurement of the intracellular glass transition temperature of a mammalian cell type under cryopreservation conditions

Authors :
David E. Gilham
Stéphanie Passot
John Morris
Julie Meneghel
Victoria Sheard
Fernanda Fonseca
Stéphanie Cenard
Génie et Microbiologie des Procédés Alimentaires (GMPA)
Institut National de la Recherche Agronomique (INRA)-AgroParisTech
Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP)
Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse)
Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)
Institute of Cancer Sciences
University of Manchester
Asymptote Ltd
AgroParisTech-Institut National de la Recherche Agronomique (INRA)
Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse)
Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA)
Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)
Source :
Cryobiology, 52nd Annual Meeting of the Society for Crryobiology, 52nd Annual Meeting of the Society for Crryobiology, Jul 2015, Ostrava, Czech Republic. pp.538, ⟨10.1016/j.cryobiol.2015.10.008⟩
Publication Year :
2015
Publisher :
Elsevier BV, 2015.

Abstract

A key physical event during the cooling of cells is the temperature at which the intracellular compartment undergoes vitrification. Recently (http://dx. doi.org/10.1371/journal.pone.0066207) we have measured the intracellular glass transition temperature (Tg) of bacteria using differential scanning calorimetry (DSC). Here we report the first data on the intracellular Tg of a mammalian cell cooled with and without a cryoprotectant (Me2SO). Jurkat cells (an immortalized T cell line) were concentrated and a pellet of approximately 30 mg cooled in a DSC, SnowMax was added to induce ice nucleation. On warming an intracellular glass transition was detected by the deviation in the first derivative of the heat flow, associated with a change in heat capacity as the cells devitrify at Tg. The transition temperature measured by DSC is Tg', that is the Tg of the maximally freezeconcentrated intracellular matrix. In the presence of Me2SO intracellular Tg was measured to be - 40 C, in controls (no Me2SO) Tg occurred at higher temperatures. In further experiments the temperature of the lipid phase transitions were analyzed by FTIR, and were measured to occur above freezing temperatures. The implications of the measured values of Tg’ for the basic cryobiology of mammalian cells will be discussed.

Details

ISSN :
00112240
Volume :
71
Database :
OpenAIRE
Journal :
Cryobiology
Accession number :
edsair.doi.dedup.....e1930e4ece4f49c21fd688cdb50b4fee
Full Text :
https://doi.org/10.1016/j.cryobiol.2015.10.008