Membrane hydration correlates to cellular biophysics during freezing in mammalian cells

Saravana K. Balasubramanian; Willem F. Wolkers; John C. Bischof

doi:10.1016/j.bbamem.2009.02.009

Membrane hydration correlates to cellular biophysics during freezing in mammalian cells

Saravana K. Balasubramanian, Willem F. Wolkers, John C. Bischof

Mechanical Engineering

Research output: Contribution to journal › Article › peer-review

50 Scopus citations

Abstract

Cell survival during freezing applications in biomedicine is highly correlated to the temperature history and its dependent cellular biophysical events of dehydration and intracellular ice formation (IIF). Although cell membranes are known to play a significant role in cell injury, a clear correlation between the membrane state and the surrounding intracellular and extracellular water is still lacking. We previously showed that lipid hydration in LNCaP tumor cells is related to cellular dehydration. The goal of this study is to build upon this work by correlating both the phase state of the membrane and the surrounding water to cellular biophysical events in three different mammalian cell types: human prostate tumor cells (LNCaP), human dermal fibroblasts (HDF), and porcine smooth muscle cells (SMC) using Fourier Transform Infrared spectroscopy (FTIR). Variable cooling rates were achieved by controlling the degree of supercooling prior to ice nucleation (- 3 °C and - 10 °C) while the sample was cooled at a set rate of 2 °C/min. Membranes displayed a highly cooperative phase transition under dehydrating conditions (i.e. NT = - 3 °C), which was not observed under IIF conditions (NT = - 10 °C). Spectral analysis showed a consistently greater amount of ice formation during dehydrating vs. IIF conditions in all cell types. This is hypothesized to be due to the extreme loss of membrane hydration in dehydrating cells that is manifested as excess water available for phase change. Interestingly, changes in residual membrane conformational disorder correlate strongly with cellular volumetric decreases as assessed by cryomicroscopy. A strong correlation was also found between the activation energies for freezing induced lyotropic membrane phase change determined using FTIR and the water transport measured by cryomicroscopy. Reduced lipid hydration under dehydration freezing conditions is suggested as one of the likely causes of what has been termed as "solution effects" injury in cryobiology.

Original language	English (US)
Pages (from-to)	945-953
Number of pages	9
Journal	Biochimica et Biophysica Acta - Biomembranes
Volume	1788
Issue number	5
DOIs	https://doi.org/10.1016/j.bbamem.2009.02.009
State	Published - May 2009

Bibliographical note

Funding Information:
The authors would like to acknowledge grant support from National Institute of Health (NIH) R01-CA07528. The authors would also like to thank Dr. Alptekin Aksan for valuable discussions, Dr. Joel Slaton and the MD Anderson Cancer Center for the LNCaP Pro5 line cells.

Keywords

Cryomicroscopy
Cryopreservation
Cryosurgery
FTIR
Membrane phase behavior

Access

10.1016/j.bbamem.2009.02.009

OpenUrl availability

Full text

Cite this

@article{01c678bffdd94b3789a8b40d639ccf7d,

title = "Membrane hydration correlates to cellular biophysics during freezing in mammalian cells",

abstract = "Cell survival during freezing applications in biomedicine is highly correlated to the temperature history and its dependent cellular biophysical events of dehydration and intracellular ice formation (IIF). Although cell membranes are known to play a significant role in cell injury, a clear correlation between the membrane state and the surrounding intracellular and extracellular water is still lacking. We previously showed that lipid hydration in LNCaP tumor cells is related to cellular dehydration. The goal of this study is to build upon this work by correlating both the phase state of the membrane and the surrounding water to cellular biophysical events in three different mammalian cell types: human prostate tumor cells (LNCaP), human dermal fibroblasts (HDF), and porcine smooth muscle cells (SMC) using Fourier Transform Infrared spectroscopy (FTIR). Variable cooling rates were achieved by controlling the degree of supercooling prior to ice nucleation (- 3 °C and - 10 °C) while the sample was cooled at a set rate of 2 °C/min. Membranes displayed a highly cooperative phase transition under dehydrating conditions (i.e. NT = - 3 °C), which was not observed under IIF conditions (NT = - 10 °C). Spectral analysis showed a consistently greater amount of ice formation during dehydrating vs. IIF conditions in all cell types. This is hypothesized to be due to the extreme loss of membrane hydration in dehydrating cells that is manifested as excess water available for phase change. Interestingly, changes in residual membrane conformational disorder correlate strongly with cellular volumetric decreases as assessed by cryomicroscopy. A strong correlation was also found between the activation energies for freezing induced lyotropic membrane phase change determined using FTIR and the water transport measured by cryomicroscopy. Reduced lipid hydration under dehydration freezing conditions is suggested as one of the likely causes of what has been termed as {"}solution effects{"} injury in cryobiology.",

keywords = "Cryomicroscopy, Cryopreservation, Cryosurgery, FTIR, Membrane phase behavior",

author = "Balasubramanian, {Saravana K.} and Wolkers, {Willem F.} and Bischof, {John C.}",

note = "Funding Information: The authors would like to acknowledge grant support from National Institute of Health (NIH) R01-CA07528. The authors would also like to thank Dr. Alptekin Aksan for valuable discussions, Dr. Joel Slaton and the MD Anderson Cancer Center for the LNCaP Pro5 line cells.",

year = "2009",

month = may,

doi = "10.1016/j.bbamem.2009.02.009",

language = "English (US)",

volume = "1788",

pages = "945--953",

journal = "Biochimica et Biophysica Acta - Biomembranes",

issn = "0005-2736",

publisher = "Elsevier",

number = "5",

}

TY - JOUR

T1 - Membrane hydration correlates to cellular biophysics during freezing in mammalian cells

AU - Balasubramanian, Saravana K.

AU - Wolkers, Willem F.

AU - Bischof, John C.

N1 - Funding Information: The authors would like to acknowledge grant support from National Institute of Health (NIH) R01-CA07528. The authors would also like to thank Dr. Alptekin Aksan for valuable discussions, Dr. Joel Slaton and the MD Anderson Cancer Center for the LNCaP Pro5 line cells.

PY - 2009/5

Y1 - 2009/5

N2 - Cell survival during freezing applications in biomedicine is highly correlated to the temperature history and its dependent cellular biophysical events of dehydration and intracellular ice formation (IIF). Although cell membranes are known to play a significant role in cell injury, a clear correlation between the membrane state and the surrounding intracellular and extracellular water is still lacking. We previously showed that lipid hydration in LNCaP tumor cells is related to cellular dehydration. The goal of this study is to build upon this work by correlating both the phase state of the membrane and the surrounding water to cellular biophysical events in three different mammalian cell types: human prostate tumor cells (LNCaP), human dermal fibroblasts (HDF), and porcine smooth muscle cells (SMC) using Fourier Transform Infrared spectroscopy (FTIR). Variable cooling rates were achieved by controlling the degree of supercooling prior to ice nucleation (- 3 °C and - 10 °C) while the sample was cooled at a set rate of 2 °C/min. Membranes displayed a highly cooperative phase transition under dehydrating conditions (i.e. NT = - 3 °C), which was not observed under IIF conditions (NT = - 10 °C). Spectral analysis showed a consistently greater amount of ice formation during dehydrating vs. IIF conditions in all cell types. This is hypothesized to be due to the extreme loss of membrane hydration in dehydrating cells that is manifested as excess water available for phase change. Interestingly, changes in residual membrane conformational disorder correlate strongly with cellular volumetric decreases as assessed by cryomicroscopy. A strong correlation was also found between the activation energies for freezing induced lyotropic membrane phase change determined using FTIR and the water transport measured by cryomicroscopy. Reduced lipid hydration under dehydration freezing conditions is suggested as one of the likely causes of what has been termed as "solution effects" injury in cryobiology.

AB - Cell survival during freezing applications in biomedicine is highly correlated to the temperature history and its dependent cellular biophysical events of dehydration and intracellular ice formation (IIF). Although cell membranes are known to play a significant role in cell injury, a clear correlation between the membrane state and the surrounding intracellular and extracellular water is still lacking. We previously showed that lipid hydration in LNCaP tumor cells is related to cellular dehydration. The goal of this study is to build upon this work by correlating both the phase state of the membrane and the surrounding water to cellular biophysical events in three different mammalian cell types: human prostate tumor cells (LNCaP), human dermal fibroblasts (HDF), and porcine smooth muscle cells (SMC) using Fourier Transform Infrared spectroscopy (FTIR). Variable cooling rates were achieved by controlling the degree of supercooling prior to ice nucleation (- 3 °C and - 10 °C) while the sample was cooled at a set rate of 2 °C/min. Membranes displayed a highly cooperative phase transition under dehydrating conditions (i.e. NT = - 3 °C), which was not observed under IIF conditions (NT = - 10 °C). Spectral analysis showed a consistently greater amount of ice formation during dehydrating vs. IIF conditions in all cell types. This is hypothesized to be due to the extreme loss of membrane hydration in dehydrating cells that is manifested as excess water available for phase change. Interestingly, changes in residual membrane conformational disorder correlate strongly with cellular volumetric decreases as assessed by cryomicroscopy. A strong correlation was also found between the activation energies for freezing induced lyotropic membrane phase change determined using FTIR and the water transport measured by cryomicroscopy. Reduced lipid hydration under dehydration freezing conditions is suggested as one of the likely causes of what has been termed as "solution effects" injury in cryobiology.

KW - Cryomicroscopy

KW - Cryopreservation

KW - Cryosurgery

KW - FTIR

KW - Membrane phase behavior

UR - http://www.scopus.com/inward/record.url?scp=65249190875&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=65249190875&partnerID=8YFLogxK

U2 - 10.1016/j.bbamem.2009.02.009

DO - 10.1016/j.bbamem.2009.02.009

M3 - Article

C2 - 19233120

AN - SCOPUS:65249190875

SN - 0005-2736

VL - 1788

SP - 945

EP - 953

JO - Biochimica et Biophysica Acta - Biomembranes

JF - Biochimica et Biophysica Acta - Biomembranes

IS - 5

ER -

Membrane hydration correlates to cellular biophysics during freezing in mammalian cells

Abstract

Bibliographical note

Keywords

Access

OpenUrl availability

Other files and links

Fingerprint

Cite this