Cryopreservation of cells: FT-IR monitoring of lipid membrane at freeze-thaw cycles.
In the present study, FTIR spectroscopy was used to monitor the freeze-thaw cycle of two cellular lines (HuDe and Jurkat) suspended in three different media: phosphate buffer solution (PBS); dimethylsulfoxide (DMSO)/PBS solution at 0.1 DMSO molar fraction; and CryoSure (0.1 DMSO molar fraction PBS solution+dextran 5% w/v) solution. The Trypan Blue test was also applied before freezing and after thawing each cell sample to estimate the recovery of membrane integrity after thermal treatment, and correlate this datum with spectroscopic results. By following the temperature evolution of two different spectral components (the libration and bending combination mode νc(H2O) at 2000-2500 cm(-1), and the methylene symmetric stretching vibration νsym(CH2) at about 2850 cm(-1)) in the -120÷28°C range, we evidenced the main transition of lipid membrane in connection with cell dehydration, as induced by ice formation in the extracellular medium. In particular, in DMSO/PBS and CryoSure samples we observed a transition to a more rigid state of the lipid membrane together with an increased amount of non-freezable water in the extracellular medium; these results are connected to the role of DMSO as a cryoprotective agent irrespective of the nature of cell type.
Dipartimento di Chimica, Biologia e Biotecnologie, Università di Perugia, Via Elce di sotto 8, 06123 Perugia, Italy.
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Giugliarelli
Dipartimento di Chimica, Biologia e Biotecnologie, Università di Perugia, Via Elce di sotto 8, 06123 Perugia, Italy. Electronic address: paola.sassi@unipg.it.
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Sassi
Dipartimento di Chimica, Biologia e Biotecnologie, Università di Perugia, Via Elce di sotto 8, 06123 Perugia, Italy.
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Urbanelli
Dipartimento di Chimica, Biologia e Biotecnologie, Università di Perugia, Via Elce di sotto 8, 06123 Perugia, Italy.
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Paolantoni
Istituto Officina dei Materiali del CNR (CNR-IOM) - Unità di Perugia, c/o Dipartimento di Fisica e Geologia, Via Pascoli, Perugia, Italy.
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Caponi
Dipartimento di Chimica, Biologia e Biotecnologie, Università di Perugia, Via Elce di sotto 8, 06123 Perugia, Italy; Istituto di Biofisica del CNR (CNR-IBF) - Unità di Trento, c/o Fondazione Bruno Kessler, Via alla Cascata 56/C, 38123 Trento, Italy.
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Ricci
Dipartimento di Chimica, Biologia e Biotecnologie, Università di Perugia, Via Elce di sotto 8, 06123 Perugia, Italy; Centro di Eccellenza sui Materiali Innovativi Nanostrutturati (CEMIN), Università di Perugia, Italy.
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Emiliani
Centro di Eccellenza sui Materiali Innovativi Nanostrutturati (CEMIN), Università di Perugia, Italy; Dipartimento di Fisica e Geologia, Via Pascoli, Università di Perugia, Italy.
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Fioretto
Dipartimento di Chimica, Biologia e Biotecnologie, Università di Perugia, Via Elce di sotto 8, 06123 Perugia, Italy.
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Morresi
D002021:Buffers; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry*; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000737:chemistry*; D003911:Dextrans / Q000737:chemistry; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D005347:Fibroblasts / Q000166:cytology*; D005615:Freezing; D006801:Humans; D019169:Jurkat Cells; D008563:Membrane Lipids / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature
D002021:Buffers; D003451:Cryoprotective Agents; D003911:Dextrans; D008563:Membrane Lipids; D010710:Phosphates; D004121:Dimethyl Sulfoxide
Cell; Cryopreservation; Dimethylsulfoxide; FTIR spectroscopy; Lipid phase behavior