Evaluation of Different Cryoprotectant Combinations in Vitrification and Slow Freezing for Ovarian Tissue Preservation in Domestic Cats
ABSTRACT Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising this, it becomes imperative to implement strategies aimed at mitigating this concerning conservation scenario. For this, female fertil...
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| Published in | Reproduction in domestic animals Vol. 60; no. 4; pp. e70064 - n/a |
|---|---|
| Main Authors | , , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Germany
Blackwell Publishing Ltd
01.04.2025
John Wiley and Sons Inc |
| Subjects | |
| Online Access | Get full text |
| ISSN | 0936-6768 1439-0531 1439-0531 |
| DOI | 10.1111/rda.70064 |
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| Abstract | ABSTRACT
Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising this, it becomes imperative to implement strategies aimed at mitigating this concerning conservation scenario. For this, female fertility preservation is crucial in this context, and studies concerning this field are still scarce. In the realm of cryopreservation, prevalent methods involve slow freezing (SF) and vitrification (V). This study aimed to evaluate various cryoprotective combinations for V or SF processes applied to domestic cat ovarian tissue. Twenty ovaries from 10 healthy cats were dissected, and cortical regions were sectioned into eight fragments measuring 3 mm3 each. These fragments were randomly allocated to three different treatment groups for V (V1, V2 and V3) or SF (SF1, SF2 and SF3). Each group employed solutions with varying concentrations of DMSO, EG and either trehalose or sucrose. The assessment included histological evaluation, follicle counting, immunohistochemical analysis of proliferative activity, and ultrastructural examination. The results demonstrated that the V1 protocol—composed of an equilibration solution with 10% DMSO, 10% EG and 0.1 M trehalose, followed by a V solution with 20% DMSO, 20% EG and 0.1 M trehalose—proved most effective. This combination best preserved follicular morphology, reduced degeneration, supported follicle proliferation and maintained favourable ultrastructural integrity compared to other treatments. These findings provide a valuable foundation for improving fertility preservation in domestic cats, with potential applications for endangered felid conservation programs. |
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| AbstractList | ABSTRACT
Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising this, it becomes imperative to implement strategies aimed at mitigating this concerning conservation scenario. For this, female fertility preservation is crucial in this context, and studies concerning this field are still scarce. In the realm of cryopreservation, prevalent methods involve slow freezing (SF) and vitrification (V). This study aimed to evaluate various cryoprotective combinations for V or SF processes applied to domestic cat ovarian tissue. Twenty ovaries from 10 healthy cats were dissected, and cortical regions were sectioned into eight fragments measuring 3 mm3 each. These fragments were randomly allocated to three different treatment groups for V (V1, V2 and V3) or SF (SF1, SF2 and SF3). Each group employed solutions with varying concentrations of DMSO, EG and either trehalose or sucrose. The assessment included histological evaluation, follicle counting, immunohistochemical analysis of proliferative activity, and ultrastructural examination. The results demonstrated that the V1 protocol—composed of an equilibration solution with 10% DMSO, 10% EG and 0.1 M trehalose, followed by a V solution with 20% DMSO, 20% EG and 0.1 M trehalose—proved most effective. This combination best preserved follicular morphology, reduced degeneration, supported follicle proliferation and maintained favourable ultrastructural integrity compared to other treatments. These findings provide a valuable foundation for improving fertility preservation in domestic cats, with potential applications for endangered felid conservation programs. Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising this, it becomes imperative to implement strategies aimed at mitigating this concerning conservation scenario. For this, female fertility preservation is crucial in this context, and studies concerning this field are still scarce. In the realm of cryopreservation, prevalent methods involve slow freezing (SF) and vitrification (V). This study aimed to evaluate various cryoprotective combinations for V or SF processes applied to domestic cat ovarian tissue. Twenty ovaries from 10 healthy cats were dissected, and cortical regions were sectioned into eight fragments measuring 3 mm3 each. These fragments were randomly allocated to three different treatment groups for V (V1, V2 and V3) or SF (SF1, SF2 and SF3). Each group employed solutions with varying concentrations of DMSO, EG and either trehalose or sucrose. The assessment included histological evaluation, follicle counting, immunohistochemical analysis of proliferative activity, and ultrastructural examination. The results demonstrated that the V1 protocol—composed of an equilibration solution with 10% DMSO, 10% EG and 0.1 M trehalose, followed by a V solution with 20% DMSO, 20% EG and 0.1 M trehalose—proved most effective. This combination best preserved follicular morphology, reduced degeneration, supported follicle proliferation and maintained favourable ultrastructural integrity compared to other treatments. These findings provide a valuable foundation for improving fertility preservation in domestic cats, with potential applications for endangered felid conservation programs. Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising this, it becomes imperative to implement strategies aimed at mitigating this concerning conservation scenario. For this, female fertility preservation is crucial in this context, and studies concerning this field are still scarce. In the realm of cryopreservation, prevalent methods involve slow freezing (SF) and vitrification (V). This study aimed to evaluate various cryoprotective combinations for V or SF processes applied to domestic cat ovarian tissue. Twenty ovaries from 10 healthy cats were dissected, and cortical regions were sectioned into eight fragments measuring 3 mm 3 each. These fragments were randomly allocated to three different treatment groups for V (V1, V2 and V3) or SF (SF1, SF2 and SF3). Each group employed solutions with varying concentrations of DMSO, EG and either trehalose or sucrose. The assessment included histological evaluation, follicle counting, immunohistochemical analysis of proliferative activity, and ultrastructural examination. The results demonstrated that the V1 protocol—composed of an equilibration solution with 10% DMSO, 10% EG and 0.1 M trehalose, followed by a V solution with 20% DMSO, 20% EG and 0.1 M trehalose—proved most effective. This combination best preserved follicular morphology, reduced degeneration, supported follicle proliferation and maintained favourable ultrastructural integrity compared to other treatments. These findings provide a valuable foundation for improving fertility preservation in domestic cats, with potential applications for endangered felid conservation programs. Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising this, it becomes imperative to implement strategies aimed at mitigating this concerning conservation scenario. For this, female fertility preservation is crucial in this context, and studies concerning this field are still scarce. In the realm of cryopreservation, prevalent methods involve slow freezing (SF) and vitrification (V). This study aimed to evaluate various cryoprotective combinations for V or SF processes applied to domestic cat ovarian tissue. Twenty ovaries from 10 healthy cats were dissected, and cortical regions were sectioned into eight fragments measuring 3 mm 3 each. These fragments were randomly allocated to three different treatment groups for V (V1, V2 and V3) or SF (SF1, SF2 and SF3). Each group employed solutions with varying concentrations of DMSO, EG and either trehalose or sucrose. The assessment included histological evaluation, follicle counting, immunohistochemical analysis of proliferative activity, and ultrastructural examination. The results demonstrated that the V1 protocol—composed of an equilibration solution with 10% DMSO, 10% EG and 0.1 M trehalose, followed by a V solution with 20% DMSO, 20% EG and 0.1 M trehalose—proved most effective. This combination best preserved follicular morphology, reduced degeneration, supported follicle proliferation and maintained favourable ultrastructural integrity compared to other treatments. These findings provide a valuable foundation for improving fertility preservation in domestic cats, with potential applications for endangered felid conservation programs. Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising this, it becomes imperative to implement strategies aimed at mitigating this concerning conservation scenario. For this, female fertility preservation is crucial in this context, and studies concerning this field are still scarce. In the realm of cryopreservation, prevalent methods involve slow freezing (SF) and vitrification (V). This study aimed to evaluate various cryoprotective combinations for V or SF processes applied to domestic cat ovarian tissue. Twenty ovaries from 10 healthy cats were dissected, and cortical regions were sectioned into eight fragments measuring 3 mm3 each. These fragments were randomly allocated to three different treatment groups for V (V1, V2 and V3) or SF (SF1, SF2 and SF3). Each group employed solutions with varying concentrations of DMSO, EG and either trehalose or sucrose. The assessment included histological evaluation, follicle counting, immunohistochemical analysis of proliferative activity, and ultrastructural examination. The results demonstrated that the V1 protocol-composed of an equilibration solution with 10% DMSO, 10% EG and 0.1 M trehalose, followed by a V solution with 20% DMSO, 20% EG and 0.1 M trehalose-proved most effective. This combination best preserved follicular morphology, reduced degeneration, supported follicle proliferation and maintained favourable ultrastructural integrity compared to other treatments. These findings provide a valuable foundation for improving fertility preservation in domestic cats, with potential applications for endangered felid conservation programs.Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising this, it becomes imperative to implement strategies aimed at mitigating this concerning conservation scenario. For this, female fertility preservation is crucial in this context, and studies concerning this field are still scarce. In the realm of cryopreservation, prevalent methods involve slow freezing (SF) and vitrification (V). This study aimed to evaluate various cryoprotective combinations for V or SF processes applied to domestic cat ovarian tissue. Twenty ovaries from 10 healthy cats were dissected, and cortical regions were sectioned into eight fragments measuring 3 mm3 each. These fragments were randomly allocated to three different treatment groups for V (V1, V2 and V3) or SF (SF1, SF2 and SF3). Each group employed solutions with varying concentrations of DMSO, EG and either trehalose or sucrose. The assessment included histological evaluation, follicle counting, immunohistochemical analysis of proliferative activity, and ultrastructural examination. The results demonstrated that the V1 protocol-composed of an equilibration solution with 10% DMSO, 10% EG and 0.1 M trehalose, followed by a V solution with 20% DMSO, 20% EG and 0.1 M trehalose-proved most effective. This combination best preserved follicular morphology, reduced degeneration, supported follicle proliferation and maintained favourable ultrastructural integrity compared to other treatments. These findings provide a valuable foundation for improving fertility preservation in domestic cats, with potential applications for endangered felid conservation programs. Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising this, it becomes imperative to implement strategies aimed at mitigating this concerning conservation scenario. For this, female fertility preservation is crucial in this context, and studies concerning this field are still scarce. In the realm of cryopreservation, prevalent methods involve slow freezing (SF) and vitrification (V). This study aimed to evaluate various cryoprotective combinations for V or SF processes applied to domestic cat ovarian tissue. Twenty ovaries from 10 healthy cats were dissected, and cortical regions were sectioned into eight fragments measuring 3 mm each. These fragments were randomly allocated to three different treatment groups for V (V1, V2 and V3) or SF (SF1, SF2 and SF3). Each group employed solutions with varying concentrations of DMSO, EG and either trehalose or sucrose. The assessment included histological evaluation, follicle counting, immunohistochemical analysis of proliferative activity, and ultrastructural examination. The results demonstrated that the V1 protocol-composed of an equilibration solution with 10% DMSO, 10% EG and 0.1 M trehalose, followed by a V solution with 20% DMSO, 20% EG and 0.1 M trehalose-proved most effective. This combination best preserved follicular morphology, reduced degeneration, supported follicle proliferation and maintained favourable ultrastructural integrity compared to other treatments. These findings provide a valuable foundation for improving fertility preservation in domestic cats, with potential applications for endangered felid conservation programs. Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising this, it becomes imperative to implement strategies aimed at mitigating this concerning conservation scenario. For this, female fertility preservation is crucial in this context, and studies concerning this field are still scarce. In the realm of cryopreservation, prevalent methods involve slow freezing (SF) and vitrification (V). This study aimed to evaluate various cryoprotective combinations for V or SF processes applied to domestic cat ovarian tissue. Twenty ovaries from 10 healthy cats were dissected, and cortical regions were sectioned into eight fragments measuring 3 mm³ each. These fragments were randomly allocated to three different treatment groups for V (V1, V2 and V3) or SF (SF1, SF2 and SF3). Each group employed solutions with varying concentrations of DMSO, EG and either trehalose or sucrose. The assessment included histological evaluation, follicle counting, immunohistochemical analysis of proliferative activity, and ultrastructural examination. The results demonstrated that the V1 protocol—composed of an equilibration solution with 10% DMSO, 10% EG and 0.1 M trehalose, followed by a V solution with 20% DMSO, 20% EG and 0.1 M trehalose—proved most effective. This combination best preserved follicular morphology, reduced degeneration, supported follicle proliferation and maintained favourable ultrastructural integrity compared to other treatments. These findings provide a valuable foundation for improving fertility preservation in domestic cats, with potential applications for endangered felid conservation programs. |
| Author | Aguiar, Beatriz Alves Lucci, Carolina Madeira Carvalho da Costa, Marcus Vinicius Rocha Silva, Ingrid Gracielle Martins Báo, Sônia Nair Rodrigues, Aline Queiroz Ribeiro, Rayane Brandão Piau, Tathyana Benetis Santos Bezerra, Julliene Larissa Silva, Jessyca Karoline Oliveira Goulart, Jair Trapé Paulini, Fernanda Ferreira, Yasmin Barboza Bellozi, Paula Maria Quaglio |
| AuthorAffiliation | 3 Faculty of Health Sciences, Molecular Pharmacology Laboratory University of Brasilia Brasilia‐DF Brazil 2 Department of Cellular Biology University of Brasilia, Institute of Biological Sciences Brasilia‐DF Brazil 1 Department of Physiological Sciences University of Brasilia, Institute of Biological Sciences Brasilia‐DF Brazil |
| AuthorAffiliation_xml | – name: 2 Department of Cellular Biology University of Brasilia, Institute of Biological Sciences Brasilia‐DF Brazil – name: 1 Department of Physiological Sciences University of Brasilia, Institute of Biological Sciences Brasilia‐DF Brazil – name: 3 Faculty of Health Sciences, Molecular Pharmacology Laboratory University of Brasilia Brasilia‐DF Brazil |
| Author_xml | – sequence: 1 givenname: Rayane Brandão surname: Ribeiro fullname: Ribeiro, Rayane Brandão organization: University of Brasilia, Institute of Biological Sciences – sequence: 2 givenname: Aline Queiroz surname: Rodrigues fullname: Rodrigues, Aline Queiroz organization: University of Brasilia, Institute of Biological Sciences – sequence: 3 givenname: Beatriz Alves surname: Aguiar fullname: Aguiar, Beatriz Alves organization: University of Brasilia, Institute of Biological Sciences – sequence: 4 givenname: Jessyca Karoline Oliveira surname: Silva fullname: Silva, Jessyca Karoline Oliveira organization: University of Brasilia, Institute of Biological Sciences – sequence: 5 givenname: Marcus Vinicius Rocha surname: Carvalho da Costa fullname: Carvalho da Costa, Marcus Vinicius Rocha organization: University of Brasilia, Institute of Biological Sciences – sequence: 6 givenname: Julliene Larissa surname: Santos Bezerra fullname: Santos Bezerra, Julliene Larissa organization: University of Brasilia, Institute of Biological Sciences – sequence: 7 givenname: Yasmin Barboza surname: Ferreira fullname: Ferreira, Yasmin Barboza organization: University of Brasilia, Institute of Biological Sciences – sequence: 8 givenname: Ingrid Gracielle Martins surname: Silva fullname: Silva, Ingrid Gracielle Martins organization: University of Brasilia, Institute of Biological Sciences – sequence: 9 givenname: Tathyana Benetis surname: Piau fullname: Piau, Tathyana Benetis organization: University of Brasilia, Institute of Biological Sciences – sequence: 10 givenname: Carolina Madeira surname: Lucci fullname: Lucci, Carolina Madeira organization: University of Brasilia, Institute of Biological Sciences – sequence: 11 givenname: Sônia Nair surname: Báo fullname: Báo, Sônia Nair organization: University of Brasilia, Institute of Biological Sciences – sequence: 12 givenname: Jair Trapé surname: Goulart fullname: Goulart, Jair Trapé organization: University of Brasilia, Institute of Biological Sciences – sequence: 13 givenname: Paula Maria Quaglio surname: Bellozi fullname: Bellozi, Paula Maria Quaglio organization: University of Brasilia – sequence: 14 givenname: Fernanda orcidid: 0000-0003-4490-9614 surname: Paulini fullname: Paulini, Fernanda email: fepaulini@unb.br organization: University of Brasilia, Institute of Biological Sciences |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/40265625$$D View this record in MEDLINE/PubMed |
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| Copyright | 2025 The Author(s). published by Wiley‐VCH GmbH. 2025 The Author(s). Reproduction in Domestic Animals published by Wiley‐VCH GmbH. 2025. This work is published under Creative Commons Attribution – Non-Commercial – No Derivatives License~http://creativecommons.org/licenses/by-nc-nd/3.0/ (the "License"). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. |
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| Keywords | felines ovarian follicles animal reproduction cryopreservation |
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| License | Attribution-NonCommercial-NoDerivs 2025 The Author(s). Reproduction in Domestic Animals published by Wiley‐VCH GmbH. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
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| Notes | This study was financed by the Funding Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—Finance Code 001 and the University of Brasília. Rayane B. Ribeiro, Aline Q. Rodrigues and Tathyana B. Piau were supported by a National Counciil for Scientific and Technological Development (CNPq) fellowship. ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 Funding: This study was financed by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—Finance Code 001 and the University of Brasília. Rayane B. Ribeiro, Aline Q. Rodrigues and Tathyana B. Piau were supported by a National Counciil for Scientific and Technological Development (CNPq) fellowship. |
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Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family.... Over the past decade, increased hunting and habitat disturbance have significantly impacted the endangered population within the Felidae family. Recognising... |
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| SubjectTerms | animal reproduction Animals Cats Conservation Cryopreservation Cryopreservation - methods Cryopreservation - veterinary cryoprotectants Cryoprotective Agents - pharmacology Cryoprotectors Degeneration Dimethyl Sulfoxide - pharmacology domain Domestic animals Endangered populations Evaluation family Felidae felines Female female fertility Fertility Fertility Preservation - methods Fertility Preservation - veterinary Fragments Freezing habitat destruction immunohistochemistry Original Ovarian Follicle ovarian follicles Ovaries Ovary - drug effects Sucrose Trehalose Trehalose - pharmacology Vitrification |
| Title | Evaluation of Different Cryoprotectant Combinations in Vitrification and Slow Freezing for Ovarian Tissue Preservation in Domestic Cats |
| URI | https://onlinelibrary.wiley.com/doi/abs/10.1111%2Frda.70064 https://www.ncbi.nlm.nih.gov/pubmed/40265625 https://www.proquest.com/docview/3194458132 https://www.proquest.com/docview/3193718233 https://www.proquest.com/docview/3206183968 https://pubmed.ncbi.nlm.nih.gov/PMC12016461 |
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