Suprazero cooling conditions significantly influence subzero permeability parameters of mammalian ovarian tissue

R. V. Devireddy, Guohong Li, S. P. Leibo

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

To model the cryobiological responses of cells and tissues, permeability characteristics are often measured at suprazero temperatures and the measured values are used to predict the responses at subzero temperatures. The purpose of the present study was to determine whether the rate of cooling from +25 to +4°C influenced the measured water transport response of ovarian tissue at subzero temperatures in the presence or absence of cryoprotective agents (CPAs). Sections of freshly collected equine ovarian tissue were first cooled either at 40°C/min or at 0.5°C/min from 25 to 4°C, and then cooled to subzero temperatures. A shape-independent differential scanning calorimeter (DSC) technique was used to measure the volumetric shrinkage during freezing of equine ovarian tissue sections. After ice was induced to form in the extracellular fluid within the specimen, the sample was frozen from the phase change temperature to -50°C at 5°C/min. Replicate samples were frozen in isotonic medium alone or in medium containing 0.85 M glycerol or 0.85 M dimethylsulfoxide. The water transport response of ovarian tissue samples cooled at 40°C/min from 25 to 4°C was significantly different (confidence level >95%) from that of tissue samples cooled at 0.5°C/min, whether in the presence or absence of CPAs. We fitted a model of water transport to the experimentally-derived volumetric shrinkage data and determined the best-fit membrane permeability parameters (Lpg and ELp) of equine ovarian tissue during freezing. Subzero water transport parameters of ovarian tissue samples cooled at 0.5°C/min from 25 to 4°C ranged from: L pg = 0.06 to 0.73 μm /min·atm and ELp = 6.1 to 20.5 kcal/mol. The corresponding parameters of samples cooled at 40°C/min from 25 to 4°C ranged from: Lpg = 0.04 to 0.61 μm/min·atm and ELp = 8.2 to 54.2 kcal/mol. Calculations made of the theoretical response of tissue at subzero temperatures suggest that the optimal cooling rates to cryopreserve ovarian tissue are significantly dependent upon suprazero cooling conditions.

Original languageEnglish (US)
Pages (from-to)330-341
Number of pages12
JournalMolecular Reproduction and Development
Volume73
Issue number3
DOIs
StatePublished - Mar 1 2006

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Permeability
Temperature
Cryoprotective Agents
Horses
Water
Freezing
Extracellular Fluid
Ice
Dimethyl Sulfoxide
Glycerol
Membranes

All Science Journal Classification (ASJC) codes

  • Genetics
  • Developmental Biology
  • Cell Biology

Cite this

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title = "Suprazero cooling conditions significantly influence subzero permeability parameters of mammalian ovarian tissue",
abstract = "To model the cryobiological responses of cells and tissues, permeability characteristics are often measured at suprazero temperatures and the measured values are used to predict the responses at subzero temperatures. The purpose of the present study was to determine whether the rate of cooling from +25 to +4°C influenced the measured water transport response of ovarian tissue at subzero temperatures in the presence or absence of cryoprotective agents (CPAs). Sections of freshly collected equine ovarian tissue were first cooled either at 40°C/min or at 0.5°C/min from 25 to 4°C, and then cooled to subzero temperatures. A shape-independent differential scanning calorimeter (DSC) technique was used to measure the volumetric shrinkage during freezing of equine ovarian tissue sections. After ice was induced to form in the extracellular fluid within the specimen, the sample was frozen from the phase change temperature to -50°C at 5°C/min. Replicate samples were frozen in isotonic medium alone or in medium containing 0.85 M glycerol or 0.85 M dimethylsulfoxide. The water transport response of ovarian tissue samples cooled at 40°C/min from 25 to 4°C was significantly different (confidence level >95{\%}) from that of tissue samples cooled at 0.5°C/min, whether in the presence or absence of CPAs. We fitted a model of water transport to the experimentally-derived volumetric shrinkage data and determined the best-fit membrane permeability parameters (Lpg and ELp) of equine ovarian tissue during freezing. Subzero water transport parameters of ovarian tissue samples cooled at 0.5°C/min from 25 to 4°C ranged from: L pg = 0.06 to 0.73 μm /min·atm and ELp = 6.1 to 20.5 kcal/mol. The corresponding parameters of samples cooled at 40°C/min from 25 to 4°C ranged from: Lpg = 0.04 to 0.61 μm/min·atm and ELp = 8.2 to 54.2 kcal/mol. Calculations made of the theoretical response of tissue at subzero temperatures suggest that the optimal cooling rates to cryopreserve ovarian tissue are significantly dependent upon suprazero cooling conditions.",
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Suprazero cooling conditions significantly influence subzero permeability parameters of mammalian ovarian tissue. / Devireddy, R. V.; Li, Guohong; Leibo, S. P.

In: Molecular Reproduction and Development, Vol. 73, No. 3, 01.03.2006, p. 330-341.

Research output: Contribution to journalArticle

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AB - To model the cryobiological responses of cells and tissues, permeability characteristics are often measured at suprazero temperatures and the measured values are used to predict the responses at subzero temperatures. The purpose of the present study was to determine whether the rate of cooling from +25 to +4°C influenced the measured water transport response of ovarian tissue at subzero temperatures in the presence or absence of cryoprotective agents (CPAs). Sections of freshly collected equine ovarian tissue were first cooled either at 40°C/min or at 0.5°C/min from 25 to 4°C, and then cooled to subzero temperatures. A shape-independent differential scanning calorimeter (DSC) technique was used to measure the volumetric shrinkage during freezing of equine ovarian tissue sections. After ice was induced to form in the extracellular fluid within the specimen, the sample was frozen from the phase change temperature to -50°C at 5°C/min. Replicate samples were frozen in isotonic medium alone or in medium containing 0.85 M glycerol or 0.85 M dimethylsulfoxide. The water transport response of ovarian tissue samples cooled at 40°C/min from 25 to 4°C was significantly different (confidence level >95%) from that of tissue samples cooled at 0.5°C/min, whether in the presence or absence of CPAs. We fitted a model of water transport to the experimentally-derived volumetric shrinkage data and determined the best-fit membrane permeability parameters (Lpg and ELp) of equine ovarian tissue during freezing. Subzero water transport parameters of ovarian tissue samples cooled at 0.5°C/min from 25 to 4°C ranged from: L pg = 0.06 to 0.73 μm /min·atm and ELp = 6.1 to 20.5 kcal/mol. The corresponding parameters of samples cooled at 40°C/min from 25 to 4°C ranged from: Lpg = 0.04 to 0.61 μm/min·atm and ELp = 8.2 to 54.2 kcal/mol. Calculations made of the theoretical response of tissue at subzero temperatures suggest that the optimal cooling rates to cryopreserve ovarian tissue are significantly dependent upon suprazero cooling conditions.

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