Positive impact of sucrose supplementation during slow freezing of cat ovarian tissues on cellular viability, follicle morphology, and DNA integrity

The objectives of the study were to (1) examine and optimize the impact of sucrose during slow freezing and (2) compare the results of two freezing methods (slow freezing and vitrification) on cellular viability (germinal and stromal cells), follicle morphology, DNA integrity, and gap junction prote...

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Published in:Theriogenology 2015-06, Vol.83 (9), p.1553-1561
Main Authors: Tanpradit, Nae, Comizzoli, Pierre, Srisuwatanasagul, Sayamon, Chatdarong, Kaywalee
Format: Article
Language:English
Subjects:
Animals
Apoptosis - drug effects
Cats
Cell Survival
Connexin 43
Cryopreservation
Cryopreservation - methods
Cryopreservation - veterinary
DNA Damage
Domestic cat
Female
Ovarian Follicle - anatomy & histology
Ovarian Follicle - drug effects
Ovarian Follicle - physiology
Ovarian tissue
Ovary - physiology
Sucrose
Sucrose - pharmacology
Vitrification
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cited_by cdi_FETCH-LOGICAL-c456t-b6c940d325f08a80dd8744bcba8778c672ac449da5ff983dbf72562bcad368cc3
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container_issue 9
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creator Tanpradit, Nae
Comizzoli, Pierre
Srisuwatanasagul, Sayamon
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description The objectives of the study were to (1) examine and optimize the impact of sucrose during slow freezing and (2) compare the results of two freezing methods (slow freezing and vitrification) on cellular viability (germinal and stromal cells), follicle morphology, DNA integrity, and gap junction protein expression (connexin 43 [Cx 43]). Different sucrose supplementations (0, 0.1, and 0.3 M) in standard freezing medium were compared before and after slow freezing. Ovarian tissue slow frozen using 0.1- (4.0 ± 0.4) or 0.3-M sucrose (3.9 ± 0.5) yielded better follicular viability (number of positive follicles per 0.0625 mm2) than the group without sucrose (1.9 ± 0.2; P 
doi_str_mv 10.1016/j.theriogenology.2015.01.035
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Different sucrose supplementations (0, 0.1, and 0.3 M) in standard freezing medium were compared before and after slow freezing. Ovarian tissue slow frozen using 0.1- (4.0 ± 0.4) or 0.3-M sucrose (3.9 ± 0.5) yielded better follicular viability (number of positive follicles per 0.0625 mm2) than the group without sucrose (1.9 ± 0.2; P < 0.05). Morphologically normal primordial follicles were higher in the sucrose-treated groups (0.1 M, 47.4% and 0.3 M, 43.5%) than the group without sucrose (0 M, 33.8%; P < 0.05). Moreover, less apoptotic primordial follicles were found in both sucrose groups (0.1 M, 1.2% and 0.3 M, 1.9%) than the group without sucrose (7.7%; P < 0.05). However, their Cx 43 expression showed no difference among the groups of different sucrose concentrations. In terms of the freezing methods used, vitrified ovarian tissues had fewer viable follicles (3.2 ± 0.6) than the slow-freezing method (4.6 ± 0.6; P < 0.05). In addition, the slow freezing resulted in more postthawed morphologically normal primordial follicles (38.8% vs. 28.3%, P < 0.05) and less apoptotic primordial follicles (3.8% vs. 8.9%, P < 0.05) than vitrification. The Cx 43 expression showed no difference between slow freezing and vitrification. The present study reported the positive effects of sucrose supplementation and slow-freezing method on the follicular viability, follicular histologic appearances of follicles, and apoptosis of the follicles and stromal cells in cat ovarian tissues.]]></description><identifier>ISSN: 0093-691X</identifier><identifier>EISSN: 1879-3231</identifier><identifier>DOI: 10.1016/j.theriogenology.2015.01.035</identifier><identifier>PMID: 25747194</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Apoptosis - drug effects ; Cats ; Cell Survival ; Connexin 43 ; Cryopreservation ; Cryopreservation - methods ; Cryopreservation - veterinary ; DNA Damage ; Domestic cat ; Female ; Ovarian Follicle - anatomy &amp; histology ; Ovarian Follicle - drug effects ; Ovarian Follicle - physiology ; Ovarian tissue ; Ovary - physiology ; Sucrose ; Sucrose - pharmacology ; Vitrification</subject><ispartof>Theriogenology, 2015-06, Vol.83 (9), p.1553-1561</ispartof><rights>2015 Elsevier Inc.</rights><rights>Copyright © 2015 Elsevier Inc. 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Different sucrose supplementations (0, 0.1, and 0.3 M) in standard freezing medium were compared before and after slow freezing. Ovarian tissue slow frozen using 0.1- (4.0 ± 0.4) or 0.3-M sucrose (3.9 ± 0.5) yielded better follicular viability (number of positive follicles per 0.0625 mm2) than the group without sucrose (1.9 ± 0.2; P < 0.05). Morphologically normal primordial follicles were higher in the sucrose-treated groups (0.1 M, 47.4% and 0.3 M, 43.5%) than the group without sucrose (0 M, 33.8%; P < 0.05). Moreover, less apoptotic primordial follicles were found in both sucrose groups (0.1 M, 1.2% and 0.3 M, 1.9%) than the group without sucrose (7.7%; P < 0.05). However, their Cx 43 expression showed no difference among the groups of different sucrose concentrations. In terms of the freezing methods used, vitrified ovarian tissues had fewer viable follicles (3.2 ± 0.6) than the slow-freezing method (4.6 ± 0.6; P < 0.05). In addition, the slow freezing resulted in more postthawed morphologically normal primordial follicles (38.8% vs. 28.3%, P < 0.05) and less apoptotic primordial follicles (3.8% vs. 8.9%, P < 0.05) than vitrification. The Cx 43 expression showed no difference between slow freezing and vitrification. 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Different sucrose supplementations (0, 0.1, and 0.3 M) in standard freezing medium were compared before and after slow freezing. Ovarian tissue slow frozen using 0.1- (4.0 ± 0.4) or 0.3-M sucrose (3.9 ± 0.5) yielded better follicular viability (number of positive follicles per 0.0625 mm2) than the group without sucrose (1.9 ± 0.2; P < 0.05). Morphologically normal primordial follicles were higher in the sucrose-treated groups (0.1 M, 47.4% and 0.3 M, 43.5%) than the group without sucrose (0 M, 33.8%; P < 0.05). Moreover, less apoptotic primordial follicles were found in both sucrose groups (0.1 M, 1.2% and 0.3 M, 1.9%) than the group without sucrose (7.7%; P < 0.05). However, their Cx 43 expression showed no difference among the groups of different sucrose concentrations. In terms of the freezing methods used, vitrified ovarian tissues had fewer viable follicles (3.2 ± 0.6) than the slow-freezing method (4.6 ± 0.6; P < 0.05). In addition, the slow freezing resulted in more postthawed morphologically normal primordial follicles (38.8% vs. 28.3%, P < 0.05) and less apoptotic primordial follicles (3.8% vs. 8.9%, P < 0.05) than vitrification. The Cx 43 expression showed no difference between slow freezing and vitrification. The present study reported the positive effects of sucrose supplementation and slow-freezing method on the follicular viability, follicular histologic appearances of follicles, and apoptosis of the follicles and stromal cells in cat ovarian tissues.]]></abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>25747194</pmid><doi>10.1016/j.theriogenology.2015.01.035</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0002-9470-8426</orcidid></addata></record>
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source ScienceDirect Freedom Collection 2022-2024
subjects Animals
Apoptosis - drug effects
Cats
Cell Survival
Connexin 43
Cryopreservation
Cryopreservation - methods
Cryopreservation - veterinary
DNA Damage
Domestic cat
Female
Ovarian Follicle - anatomy & histology
Ovarian Follicle - drug effects
Ovarian Follicle - physiology
Ovarian tissue
Ovary - physiology
Sucrose
Sucrose - pharmacology
Vitrification
title Positive impact of sucrose supplementation during slow freezing of cat ovarian tissues on cellular viability, follicle morphology, and DNA integrity
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