The rate at which equine and macaque ovarian tissue sections are first cooled from +25°Cto+4°C has a significant effect on the measured water transport when the tissues are subsequently frozen in 0.85M solutions of glycerol, dimethylsulfoxide (DMSO), or ethylene glycol (EG). To determine whether the response of ovarian tissues is altered if they are suspended in mixtures of cryoprotective agents (CPAs), rather than in solutions of a single CPA, we have now measured the subzero water transport from ovarian tissues that were suspended in mixtures of DMSO and EG. Sections of freshly collected equine and macaque ovaries were suspended either in a mixture of 0.9M EG plus 0.7M DMSO (equivalent to a mixture of 5%vv of EG and DMSO) or in a 1.6M solution of only DMSO or only EG. The tissue sections were cooled from +25°Cto+4°C and then frozen to subzero temperatures at 5°Cmin. As the tissues were being frozen, a shape-independent differential scanning calorimeter technique was used to measure water loss from the tissues and, consequently, the best fit membrane permeability parameters (Lpg and ELp) of ovarian tissues during freezing. In the mixture of DMSO+EG, the respective values of Lpg and ELp for equine tissue first cooled at 40°Cmin between +25°C and +4°C before being frozen were 0.15μmminatm and 7.6kcalmole. The corresponding Lpg and ELp values for equine tissue suspended in 1.6M DMSO were 0.12μmminatm and 27.2kcalmole; in 1.6M EG, the values were 0.06μmminatm and 21.9kcalmole, respectively. For macaque ovarian tissues suspended in the mixture of DMSO+EG, the respective values of Lpg and ELp were 0.26μmminatm and 26.2kcalmole. Similarly, the corresponding LLg and ELp values for macaque tissue suspended in 1.6M DMSO were 0.22μmminatm and 31.4kcalmole; in 1.6M EG, the values were 0.20μmminatm and 27.9kcalmole. The parameters for both equine and macaque tissue samples suspended in the DMSO+EG mixture and first cooled at 0.5°Cmin between +25°C and +4°C were very similar to the corresponding values for samples cooled at 40°Cmin. In contrast, the membrane parameters of equine and macaque samples first cooled at 0.5°Cmin in single-component solutions were significantly different from the corresponding values for samples cooled at 40°Cmin. These results show that the membrane properties of ovarian cells from two species are different, and that the membrane properties are significantly affected both by the solution in which the tissue is suspended and by the rate at which the tissue is cooled from +25°Cto+4°C before being frozen. These observations suggest that these variables ought to be considered in the derivation of methods to cryopreserve ovarian tissues.

1.
Tulandi
,
T.
, and
Gosden
,
R.
, 2004,
Preservation of Fertility
,
Taylor & Francis
,
London, UK
.
2.
Picton
,
H. M.
,
Gosden
,
R. G.
, and
Leibo
,
S. P.
, 2002, “
Cryopreservation of Oocytes and Ovarian Tissue
,”
Current Practice and Controversies in Assisted Reproduction
,
E.
Vayena
,
P.
Rowe
, and
P. D.
Griffin
, eds.,
World Health Organization
,
Geneva
, pp.
142
151
.
3.
Parrott
,
D. M. V.
, 1960, “
The Fertility of Mice With Orthotropic Ovarian Grafts Derived From Frozen Tissue
,”
J. Reprod. Fertil.
0022-4251,
1
, pp.
230
241
.
4.
Carroll
,
J.
,
Whittingham
,
D. G.
,
Wood
,
M. J.
,
Telfer
,
E.
, and
Gosden
,
R. G.
, 1990, “
Extra-Ovarian Production of Mature Viable Mouse Oocytes From Frozen Primary Follicles
,”
J. Reprod. Fertil.
0022-4251,
90
, pp.
321
327
.
5.
Carroll
,
J.
, and
Gosden
,
R.
, 1993, “
Transplantation of Frozen-Thawed Mouse Primordial Follicles
,”
Hum. Reprod.
0268-1161,
8
, pp.
1163
1167
.
6.
Harp
,
R.
,
Leibach
,
J.
,
Black
,
J.
,
Keldahl
,
C.
, and
Karow
,
A.
, 1994, “
Cryopreservation of Murine Ovarian Tissue
,”
Cryobiology
0011-2240,
31
, pp.
336
343
.
7.
Yin
,
H.
,
Wang
,
X.
,
Kim
,
S. S.
,
Chen
,
H.
,
Tan
,
S. L.
, and
Gosden
,
R. G.
, 2003, “
Transplantation of Intact Rat Gonads Using Vascular Anastomosis: Effects of Cryopresevation, Ischaemia and Genotype
,”
Hum. Reprod.
0268-1161,
18
, pp.
1165
1172
.
8.
Gosden
,
R.
,
Baird
,
D. T.
,
Wade
,
J. C.
, and
Webb
,
R.
, 1994, “
Restoration of Fertility to Oophorectomized Sheep by Ovarian Autografts Stored at −196°C
,”
Hum. Reprod.
0268-1161,
9
, pp.
597
603
.
9.
Baird
,
D. T.
,
Webb
,
R.
,
Campbell
,
B. K.
,
Harkness
,
L. M.
, and
Gosden
,
R. G.
, 1999, “
Long-Term Ovarian Function in Sheep After Ovariectomy and Transplantation of Autografts Stored at −196°C
,”
Endocrinology
0013-7227,
140
, pp.
462
471
.
10.
Schnorr
,
J.
,
Oehninger
,
S.
,
Toner
,
J.
,
Hsiu
,
J.
,
Lazendorf
,
S.
,
Williams
,
R.
, and
Hodgen
,
G.
, 2002. “
Functional Studies of Subcutaneous Ovarian Transplants in Non-human Primates: Steroidogenesis, Endometrial Development, Ovulation, Menstrual Patterns and Gamete Morphology
,”
Hum. Reprod.
0268-1161,
17
, pp.
612
619
.
11.
Paynter
,
S. J.
,
Cooper
,
A.
,
Fuller
,
B. J.
, and
Shaw
,
R. W.
, 1999, “
Cryopreservation of Bovine Ovarian Tissue: Structural Normality of Follicles After Thawing and Culture In Vitro
,”
Cryobiology
0011-2240,
38
, pp.
301
309
.
12.
Newton
,
H.
,
Aubard
,
Y.
,
Rutherford
,
A.
,
Sharma
,
V.
, and
Gosden
,
R. G.
, 1996, “
Low Temperature Storage and Grafting of Human Ovarian Tissue
,”
Hum. Reprod.
0268-1161,
11
, pp.
1487
1491
.
13.
Kim
,
S. S.
,
Soules
,
M. R.
, and
Battaglia
,
D. E.
, 2002, “
Follicular Development, Ovulation, and Corpus Luteum Formation in Cryopreserved Human Ovarian Tissue After Xenotransplantation
,”
Fertil. Steril.
0015-0282,
78
, pp.
77
82
.
14.
Practice Committee of the American Society for Reproductive Medicine, 2004, “
Ovarian Tissue and Oocyte Cryopreservation
,”
Fertil. Steril.
0015-0282,
82
, pp.
993
998
.
15.
Donnez
,
J.
,
Dolmans
,
M. M.
,
Demylle
,
D.
,
Jadoul
,
P.
,
Pirard
,
C.
,
Squifflet
,
J.
,
Martinez-Madrid
,
B.
, and
Van Langendonckt
,
A.
, 2004, “
Livebirth After Orthotopic Transplantation of Cryopreserved Ovarian Tissue
,”
Lancet
0140-6736,
364
, pp.
1405
1410
.
16.
Oktay
,
K.
,
Buyuk
,
E.
,
Veeck
,
L.
,
Zaninovic
,
N.
,
Xu
,
K.
,
Takeuchi
,
T.
,
Opsahl
,
M.
, and
Rosenwaks
,
Z.
, 2004, “
Embryo Development After Heterotopic Transplantation of Cryopreserved Ovarian Tissue
,”
Lancet
0140-6736,
363
, pp.
837
840
.
17.
Lobo
,
R. A.
, 2005, “
Potential Options for Preservation of Fertility in Women
,”
N. Engl. J. Med.
0028-4793,
353
, pp.
64
73
.
18.
Donnez
,
J.
,
Dolmans
,
M. M.
,
Martinez-Madrid
,
B.
,
Demylle
,
D.
, and
Van Langendonckt
,
A.
, 2005, “
The Role of Cryopreservation for Women Prior to Treatment of Malignancy
,”
Curr. Opin. Obstet. Gynecol.
1040-872X,
17
, pp.
333
338
.
19.
Devireddy
,
R. V.
, and
Bischof
,
J. C.
, 1998, “
Measurement of Water Transport during Freezing in Mammalian Liver Tissue—Part II: The Use of Differential Scanning Calorimetry
,”
ASME J. Biomech. Eng.
0148-0731,
120
, pp.
559
569
.
20.
Devireddy
,
R. V.
,
Smith
,
D. J.
, and
Bischof
,
J. C.
, 1999, “
Mass Transfer During Freezing in Rat Prostate Tumor Tissue
,”
AIChE J.
0001-1541,
45
, pp.
639
654
.
21.
Devireddy
,
R. V.
,
Coad
,
J. E.
, and
Bischof
,
J. C.
, 2001, “
Microscopic and Calorimetric Assessment of Freezing Processes in Uterine Fibroid Tissue
,”
Cryobiology
0011-2240,
42
, pp.
225
243
.
22.
Newton
,
H.
,
Fisher
,
J.
,
Arnold
,
J. R. P.
,
Pegg
,
D. E.
,
Faddy
,
M. J.
, and
Gosden
,
R. G.
, 1998. “
Permeation of Human Ovarian Tissue With Cryoprotective Agents in Preparation for Cryopreservation
,”
Hum. Reprod.
0268-1161,
13
, pp.
376
380
.
23.
Devireddy
,
R. V.
, 2005, “
Predicted Permeability Parameters of Human Ovarian Tissue Cells to Various Cryoprotectants and Water
,”
Mol. Reprod. Dev.
1040-452X,
70
, pp.
333
343
.
24.
Devireddy
,
R. V.
,
Li
,
G.
, and
Leibo
,
S. P.
, 2006, “
Suprazero Cooling Conditions Significantly Influence Subzero Permeability Parameters of Mammalian Ovarian Tissue
,”
Mol. Reprod. Dev.
1040-452X,
73
, pp.
330
341
.
25.
Li
,
G.
,
Thirumala
,
S.
,
Leibo
,
S. P.
, and
Devireddy
,
R. V.
, 2006, “
Subzero Water Transport Characteristics and Optimal Rates of Freezing Macaca mulatta (Rhesus Monkey) Ovarian Tissue
,”
Mol. Reprod. Dev.
1040-452X,
73
, pp.
1600
1611
.
26.
Yamauchi
,
Y.
,
VandeVoort
,
C. A.
, and
Leibo
,
S. P.
, 2005, “
Blastocysts Derived From Mouse Oocytes Cryopreserved by Ultra-Rapid Cooling in Low Concentrations of Cryoprotectants
,”
Cryobiology
0011-2240,
51
, p.
400
.
27.
Mazur
,
P.
, 1963, “
Kinetics of Water Loss From Cells at Subzero Temperatures and the Likelihood of Intracellular Freezing
,”
J. Gen. Physiol.
0022-1295,
47
, pp.
347
369
.
28.
Levin
,
R. L.
,
Cravalho
,
E. G.
, and
Huggins
,
C. E.
, 1976, “
A Membrane Model Describing the Effect of Temperature on the Water Conductivity of Erythrocyte Membranes at Subzero Temperatures
,”
Cryobiology
0011-2240,
13
, pp.
415
429
.
29.
Rubinsky
,
B.
, and
Pegg
,
D. E.
, 1988, “
A Mathematical Model for the Freezing Process in Biological Tissue
,”
Proc. R. Soc. London, Ser. B
0962-8452,
234
, pp.
343
358
.
30.
Smith
,
D. J.
,
Schulte
,
M.
, and
Bischof
,
J. C.
, 1998, “
The Effect of Dimethylsulfoxide on the Water Transport Response of Rat Hepatocytes During Freezing
,”
ASME J. Biomech. Eng.
0148-0731,
120
, pp.
549
558
.
31.
Mazur
,
P.
, 2004, “
Principles of Cryobiology
,”
Life in the Frozen State
,
B. J.
Fuller
,
N.
Lane
, and
E. E.
Bensos
, eds.
CRC
,
Boca Raton, FL
, pp.
3
66
.
32.
Toner
,
M.
, 1993, “
Nucleation of Ice Crystals in Biological Cells
,”
Advances in Low-Temperature Biology
,”
P. L.
Steponkus
, ed.,
JAI Press
,
London
, pp.
1
52
.
33.
Karlsson
,
J. O.
,
Cravalho
,
E. G.
, and
Toner
,
M.
, 1994, “
A Model of Diffusion-Limited Ice Growth Inside Biological Cells During Freezing
,”
J. Appl. Phys.
0021-8979,
75
, pp.
4442
4455
.
34.
Pazhayannur
,
P. V.
, and
Bischof
,
J. C.
, 1997, “
Measurement and Simulation of Water Transport During Freezing in Mammalian Liver Tissue
,”
ASME J. Biomech. Eng.
0148-0731,
119
, pp.
269
277
.
35.
Schneider
,
U.
, and
Mazur
,
P.
, 1984, “
Osmotic Consequences of Cryoprotectant Permeability and its Relation to the Survival of Frozen-Thawed Embryos
,”
Theriogenology
0093-691X,
21
, pp.
68
79
.
36.
Paynter
,
S. J.
,
Borini
,
A.
,
Bianchi
,
V.
,
De Santis
,
L.
,
Flamigni
,
C.
, and
Coticchio
,
G.
, 2005, “
Volume Changes of Mature Human Oocytes on Exposure to Cryoprotectant Solutions Used in Slow Cooling Procedures
,”
Hum. Reprod.
0268-1161,
20
, pp.
1194
1199
.
37.
Kedem
,
O.
, and
Katchalsky
,
A.
, 1958, “
Thermodynamic Analysis of the Permeability of Biological Membranes to Non-Electrolytes
,”
Biochim. Biophys. Acta
0006-3002,
27
, pp.
229
246
.
38.
Devireddy
,
R. V.
,
Raha
,
D.
, and
Bischof
,
J. C.
, 1998, “
Measurement of Water Transport During Freezing in Cell Suspensions Using a Differential Scanning Calorimeter
,”
Cryobiology
0011-2240,
36
, pp.
124
155
.
39.
Diller
,
K. R.
, 2002, “
New Techniques in Cryomicroscopy
,”
Cryobiology
0011-2240,
45
, pp.
250
251
.
40.
Yuan
,
S.
, and
Diller
,
K. R.
, 2005, “
An Optical Differential Scanning Calorimeter
,”
J. Microsc.
0022-2720
218
, pp.
85
93
.
41.
Bevington
,
P. R.
, and
Robinson
,
D. K.
, 1992,
Data Reduction and Error Analysis for the Physical Sciences
,
2nd ed.
,
McGraw-Hill
,
New York
.
42.
Thirumala
,
S.
, and
Devireddy
,
R. V.
, 2005, “
A Simplified Procedure to Determine the Optimal Rate of Freezing Biological Systems
,”
ASME J. Biomech. Eng.
0148-0731,
127
, pp.
295
300
.
43.
Dickey
,
A. N.
, and
Faller
,
R.
, 2005, “
Investigating Interactions of Biomembranes and Alcohols: A Multiscale Approach
,”
J. Polym. Sci., Part B: Polym. Phys.
0887-6266,
43
, pp.
1025
1032
.
44.
Rapatz
,
G.
, and
Luyet
,
B.
, 1968, “
Combined Effects of Freezing Rates and of Various Protective Agents on the Preservation of Human Erythrocytes
,”
Cryobiology
0011-2240,
4
, pp.
215
222
.
45.
Leibo
,
S. P.
,
Farrant
,
J.
,
Mazur
,
P.
,
Hanna
, Jr,
M. G.
, and
Smith
,
L. H.
, 1970, “
Effects of Freezing on Marrow Stem Cell Suspensions: Interactions of Cooling and Warming Rates in the Presence of PVP, Sucrose, or Glycerol
,”
Cryobiology
0011-2240,
6
, pp.
315
332
.
46.
Devireddy
,
R. V.
,
Fahrig
,
B.
,
Godke
,
R. A.
, and
Leibo
,
S. P.
, 2004, “
Subzero Water Transport Characteristics of Boar Spermatozoa Confirm Observed Optimal Cooling Rates
,”
Mol. Reprod. Dev.
1040-452X,
67
, pp.
446
457
.
47.
Thirumala
,
S.
,
Huang
,
C.
,
Dong
,
Q.
,
Tiersch
,
T. R.
, and
Devireddy
,
R. V.
, 2005, “
A Theoretically Estimated Optimal Cooling Rate for the Cryopreservation of Sperm Cells From a Live-Bearing Fish, the Green Swordtail Xiphophorus helleri
,”
Theriogenology
0093-691X,
63
, pp.
2395
2415
.
48.
Thirumala
,
S.
,
Gimble
,
J. M.
, and
Devireddy
,
R. V.
, 2005, “
Transport Phenomena During Freezing of Adipose Tissue Derived Adult Stem Cells
,”
Biotechnol. Bioeng.
0006-3592,
92
, pp.
372
383
.
49.
Devireddy
,
R. V.
,
Thirumala
,
S.
, and
Gimble
,
J. M.
, 2005, “
Cellular Response of Adipose Derived Passage-4 Adult Stem Cells to Freezing Stress
,”
ASME J. Biomech. Eng.
0148-0731,
127
, pp.
1081
1086
.
50.
Pinisetty
,
D.
,
Huang
,
C.
,
Dong
,
Q.
,
Tiersch
,
T. R.
, and
Devireddy
,
R. V.
, 2005, “
Subzero Water Permeability Parameters and Optimal Freezing Rates for Sperm Cells of the Southern Platyfish, Xiphophorus Maculatus
,”
Cryobiology
0011-2240,
50
, pp.
250
263
.
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