Details
| Original language | English |
|---|---|
| Pages (from-to) | 115-122 |
| Number of pages | 8 |
| Journal | CRYOBIOLOGY |
| Volume | 61 |
| Issue number | 1 |
| Early online date | 8 Jun 2010 |
| Publication status | Published - Aug 2010 |
Abstract
Cellular membranes are one of the primary sites of injury during freezing and thawing for cryopreservation of cells. Fourier transform infrared spectroscopy (FTIR) was used to monitor membrane phase behavior and ice formation during freezing of stallion sperm. At high subzero ice nucleation temperatures which result in cellular dehydration, membranes undergo a profound transition to a highly ordered gel phase. By contrast, low subzero nucleation temperatures, that are likely to result in intracellular ice formation, leave membrane lipids in a relatively hydrated fluid state. The extent of freezing-induced membrane dehydration was found to be dependent on the ice nucleation temperature, and showed Arrhenius behavior. The presence of glycerol did not prevent the freezing-induced membrane phase transition, but membrane dehydration occurred more gradual and over a wider temperature range. We describe a method to determine membrane hydraulic permeability parameters (ELp, Lpg) at subzero temperatures from membrane phase behavior data. In order to do this, it was assumed that the measured freezing-induced shift in wavenumber position of the symmetric CH2 stretching band arising from the lipid acyl chains is proportional to cellular dehydration. Membrane permeability parameters were also determined by analyzing the H2O-bending and -libration combination band, which yielded higher values for both ELp and Lpg as compared to lipid band analysis. These differences likely reflect differences between transport of free and membrane-bound water. FTIR allows for direct assessment of membrane properties at subzero temperatures in intact cells. The derived biophysical membrane parameters are dependent on intrinsic cell properties as well as freezing extender composition.
Keywords
- Cryobiology, Dehydration, Fourier transform infrared spectroscopy, Freezing, Membrane phase behavior, Stallion spermatozoa
ASJC Scopus subject areas
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In: CRYOBIOLOGY, Vol. 61, No. 1, 08.2010, p. 115-122.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Membrane permeability parameters for freezing of stallion sperm as determined by Fourier transform infrared spectroscopy
AU - Oldenhof, Harriëtte
AU - Friedel, Katharina
AU - Sieme, Harald
AU - Glasmacher, Birgit
AU - Wolkers, Willem F.
PY - 2010/8
Y1 - 2010/8
N2 - Cellular membranes are one of the primary sites of injury during freezing and thawing for cryopreservation of cells. Fourier transform infrared spectroscopy (FTIR) was used to monitor membrane phase behavior and ice formation during freezing of stallion sperm. At high subzero ice nucleation temperatures which result in cellular dehydration, membranes undergo a profound transition to a highly ordered gel phase. By contrast, low subzero nucleation temperatures, that are likely to result in intracellular ice formation, leave membrane lipids in a relatively hydrated fluid state. The extent of freezing-induced membrane dehydration was found to be dependent on the ice nucleation temperature, and showed Arrhenius behavior. The presence of glycerol did not prevent the freezing-induced membrane phase transition, but membrane dehydration occurred more gradual and over a wider temperature range. We describe a method to determine membrane hydraulic permeability parameters (ELp, Lpg) at subzero temperatures from membrane phase behavior data. In order to do this, it was assumed that the measured freezing-induced shift in wavenumber position of the symmetric CH2 stretching band arising from the lipid acyl chains is proportional to cellular dehydration. Membrane permeability parameters were also determined by analyzing the H2O-bending and -libration combination band, which yielded higher values for both ELp and Lpg as compared to lipid band analysis. These differences likely reflect differences between transport of free and membrane-bound water. FTIR allows for direct assessment of membrane properties at subzero temperatures in intact cells. The derived biophysical membrane parameters are dependent on intrinsic cell properties as well as freezing extender composition.
AB - Cellular membranes are one of the primary sites of injury during freezing and thawing for cryopreservation of cells. Fourier transform infrared spectroscopy (FTIR) was used to monitor membrane phase behavior and ice formation during freezing of stallion sperm. At high subzero ice nucleation temperatures which result in cellular dehydration, membranes undergo a profound transition to a highly ordered gel phase. By contrast, low subzero nucleation temperatures, that are likely to result in intracellular ice formation, leave membrane lipids in a relatively hydrated fluid state. The extent of freezing-induced membrane dehydration was found to be dependent on the ice nucleation temperature, and showed Arrhenius behavior. The presence of glycerol did not prevent the freezing-induced membrane phase transition, but membrane dehydration occurred more gradual and over a wider temperature range. We describe a method to determine membrane hydraulic permeability parameters (ELp, Lpg) at subzero temperatures from membrane phase behavior data. In order to do this, it was assumed that the measured freezing-induced shift in wavenumber position of the symmetric CH2 stretching band arising from the lipid acyl chains is proportional to cellular dehydration. Membrane permeability parameters were also determined by analyzing the H2O-bending and -libration combination band, which yielded higher values for both ELp and Lpg as compared to lipid band analysis. These differences likely reflect differences between transport of free and membrane-bound water. FTIR allows for direct assessment of membrane properties at subzero temperatures in intact cells. The derived biophysical membrane parameters are dependent on intrinsic cell properties as well as freezing extender composition.
KW - Cryobiology
KW - Dehydration
KW - Fourier transform infrared spectroscopy
KW - Freezing
KW - Membrane phase behavior
KW - Stallion spermatozoa
UR - http://www.scopus.com/inward/record.url?scp=77955419871&partnerID=8YFLogxK
U2 - 10.1016/j.cryobiol.2010.06.002
DO - 10.1016/j.cryobiol.2010.06.002
M3 - Article
C2 - 20553897
AN - SCOPUS:77955419871
VL - 61
SP - 115
EP - 122
JO - CRYOBIOLOGY
JF - CRYOBIOLOGY
SN - 0011-2240
IS - 1
ER -