Horse spermatogonial stem cell cryopreservation: feasible protocols and potential biotechnological applications
The establishment of proper conditions for spermatogonial stem cells (SSCs) cryopreservation and storage represents an important biotechnological approach for the preservation of the genetic stock of valuable animals. This study demonstrates the effects of different cryopreservation protocols on the...
| Autor principal: | Costa, Guilherme M.J. |
|---|---|
| Outros Autores: | Avelar, Gleide Fernandes, Lacerda, Samyra Maria Santos N.Nassif, Figueiredo, Andre´ Felipe Almeida, Tavares, Amanda O., Rezende-Neto, José V., Martins, Felipe G.P., França, Luiz Renato de |
| Grau: | Artigo |
| Idioma: | English |
| Publicado em: |
Cell and Tissue Research
2020
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| Acesso em linha: |
https://repositorio.inpa.gov.br/handle/1/17005 |
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oai:repositorio:1-17005 |
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oai:repositorio:1-17005 Horse spermatogonial stem cell cryopreservation: feasible protocols and potential biotechnological applications Costa, Guilherme M.J. Avelar, Gleide Fernandes Lacerda, Samyra Maria Santos N.Nassif Figueiredo, Andre´ Felipe Almeida Tavares, Amanda O. Rezende-Neto, José V. Martins, Felipe G.P. França, Luiz Renato de Dead Box4 Rna Helicase Expressed In The Germ Cells Protein Dimethyl Sulfoxide Glial Cell Line Derived Neurotrophic Factor Receptor Glial Cell Line Derived Neurotrophic Factor Receptor Alpha 1 Nanos C2hc Type Zinc Finger 2 Protein Rna Helicase Unclassified Drug Zinc Finger Protein Cryoprotective Agent Dimethyl Sulfoxide Adult Animals Cell Animals Genetics Biotechnology Cell Assay Cell Count Cell Culture Cell Function Cell Isolation Cell Structure Cell Survival Cell Viability Controlled Study Cryopreservation Fast Freezing Method Feasibility Study Genetic Gain Genetic Resources Genome Horse Immunofluorescence Test In Vitro Study Intermethod Comparison Nonhuman Phenotype Priority Journal Protein Expression Slow Freezing Method Spermatogonium Survival Rate Vitrification Adult Stem Cell Animals Cryopreservation Cytology Male Parenchyma Procedures Sperm Preservation Spermatogonium Testis Vitrification Adult Germline Stem Cells Animal Cell Survival Cryopreservation Cryoprotective Agents Dimethyl Sulfoxide Horses Male Parenchymal Tissue Semen Preservation Spermatogonia Testis Vitrification The establishment of proper conditions for spermatogonial stem cells (SSCs) cryopreservation and storage represents an important biotechnological approach for the preservation of the genetic stock of valuable animals. This study demonstrates the effects of different cryopreservation protocols on the survival rates and phenotypic expression of SSCs in horses. The cells were enzymatically isolated from testes of eight adult horses. After enrichment and characterization of germ cells in the suspension, the feasibility of several cryopreservation protocols were evaluated. Three different cryomedia compositions, associated with three different methods of freezing (vitrification, slow-freezing and fast-freezing) were evaluated. Based on the rates of viable SSCs found before and after thawing, as well as the number of recovered cells after cryopreservation, the best results were obtained utilizing the DMSO-based cryomedia associated with the slow-freezing method. In addition, when isolated cells were cultured in vitro, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and immunofluorescence analysis indicated that the cryopreserved cells were as metabolically active as the fresh cells and were also expressing typical SSCs proteins (VASA, NANOS2 and GFRA1). Therefore, our results indicate that equine SSCs can be cryopreserved without impairment of structure, function, or colony-forming abilities. © 2017, Springer-Verlag GmbH Germany. 2020-06-15T21:38:01Z 2020-06-15T21:38:01Z 2017 Artigo https://repositorio.inpa.gov.br/handle/1/17005 10.1007/s00441-017-2673-1 en Volume 370, Número 3, Pags. 489-500 Restrito Cell and Tissue Research |
| institution |
Instituto Nacional de Pesquisas da Amazônia - Repositório Institucional |
| collection |
INPA-RI |
| language |
English |
| topic |
Dead Box4 Rna Helicase Expressed In The Germ Cells Protein Dimethyl Sulfoxide Glial Cell Line Derived Neurotrophic Factor Receptor Glial Cell Line Derived Neurotrophic Factor Receptor Alpha 1 Nanos C2hc Type Zinc Finger 2 Protein Rna Helicase Unclassified Drug Zinc Finger Protein Cryoprotective Agent Dimethyl Sulfoxide Adult Animals Cell Animals Genetics Biotechnology Cell Assay Cell Count Cell Culture Cell Function Cell Isolation Cell Structure Cell Survival Cell Viability Controlled Study Cryopreservation Fast Freezing Method Feasibility Study Genetic Gain Genetic Resources Genome Horse Immunofluorescence Test In Vitro Study Intermethod Comparison Nonhuman Phenotype Priority Journal Protein Expression Slow Freezing Method Spermatogonium Survival Rate Vitrification Adult Stem Cell Animals Cryopreservation Cytology Male Parenchyma Procedures Sperm Preservation Spermatogonium Testis Vitrification Adult Germline Stem Cells Animal Cell Survival Cryopreservation Cryoprotective Agents Dimethyl Sulfoxide Horses Male Parenchymal Tissue Semen Preservation Spermatogonia Testis Vitrification |
| spellingShingle |
Dead Box4 Rna Helicase Expressed In The Germ Cells Protein Dimethyl Sulfoxide Glial Cell Line Derived Neurotrophic Factor Receptor Glial Cell Line Derived Neurotrophic Factor Receptor Alpha 1 Nanos C2hc Type Zinc Finger 2 Protein Rna Helicase Unclassified Drug Zinc Finger Protein Cryoprotective Agent Dimethyl Sulfoxide Adult Animals Cell Animals Genetics Biotechnology Cell Assay Cell Count Cell Culture Cell Function Cell Isolation Cell Structure Cell Survival Cell Viability Controlled Study Cryopreservation Fast Freezing Method Feasibility Study Genetic Gain Genetic Resources Genome Horse Immunofluorescence Test In Vitro Study Intermethod Comparison Nonhuman Phenotype Priority Journal Protein Expression Slow Freezing Method Spermatogonium Survival Rate Vitrification Adult Stem Cell Animals Cryopreservation Cytology Male Parenchyma Procedures Sperm Preservation Spermatogonium Testis Vitrification Adult Germline Stem Cells Animal Cell Survival Cryopreservation Cryoprotective Agents Dimethyl Sulfoxide Horses Male Parenchymal Tissue Semen Preservation Spermatogonia Testis Vitrification Costa, Guilherme M.J. Horse spermatogonial stem cell cryopreservation: feasible protocols and potential biotechnological applications |
| topic_facet |
Dead Box4 Rna Helicase Expressed In The Germ Cells Protein Dimethyl Sulfoxide Glial Cell Line Derived Neurotrophic Factor Receptor Glial Cell Line Derived Neurotrophic Factor Receptor Alpha 1 Nanos C2hc Type Zinc Finger 2 Protein Rna Helicase Unclassified Drug Zinc Finger Protein Cryoprotective Agent Dimethyl Sulfoxide Adult Animals Cell Animals Genetics Biotechnology Cell Assay Cell Count Cell Culture Cell Function Cell Isolation Cell Structure Cell Survival Cell Viability Controlled Study Cryopreservation Fast Freezing Method Feasibility Study Genetic Gain Genetic Resources Genome Horse Immunofluorescence Test In Vitro Study Intermethod Comparison Nonhuman Phenotype Priority Journal Protein Expression Slow Freezing Method Spermatogonium Survival Rate Vitrification Adult Stem Cell Animals Cryopreservation Cytology Male Parenchyma Procedures Sperm Preservation Spermatogonium Testis Vitrification Adult Germline Stem Cells Animal Cell Survival Cryopreservation Cryoprotective Agents Dimethyl Sulfoxide Horses Male Parenchymal Tissue Semen Preservation Spermatogonia Testis Vitrification |
| description |
The establishment of proper conditions for spermatogonial stem cells (SSCs) cryopreservation and storage represents an important biotechnological approach for the preservation of the genetic stock of valuable animals. This study demonstrates the effects of different cryopreservation protocols on the survival rates and phenotypic expression of SSCs in horses. The cells were enzymatically isolated from testes of eight adult horses. After enrichment and characterization of germ cells in the suspension, the feasibility of several cryopreservation protocols were evaluated. Three different cryomedia compositions, associated with three different methods of freezing (vitrification, slow-freezing and fast-freezing) were evaluated. Based on the rates of viable SSCs found before and after thawing, as well as the number of recovered cells after cryopreservation, the best results were obtained utilizing the DMSO-based cryomedia associated with the slow-freezing method. In addition, when isolated cells were cultured in vitro, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and immunofluorescence analysis indicated that the cryopreserved cells were as metabolically active as the fresh cells and were also expressing typical SSCs proteins (VASA, NANOS2 and GFRA1). Therefore, our results indicate that equine SSCs can be cryopreserved without impairment of structure, function, or colony-forming abilities. © 2017, Springer-Verlag GmbH Germany. |
| format |
Artigo |
| author |
Costa, Guilherme M.J. |
| author2 |
Avelar, Gleide Fernandes Lacerda, Samyra Maria Santos N.Nassif Figueiredo, Andre´ Felipe Almeida Tavares, Amanda O. Rezende-Neto, José V. Martins, Felipe G.P. França, Luiz Renato de |
| author2Str |
Avelar, Gleide Fernandes Lacerda, Samyra Maria Santos N.Nassif Figueiredo, Andre´ Felipe Almeida Tavares, Amanda O. Rezende-Neto, José V. Martins, Felipe G.P. França, Luiz Renato de |
| title |
Horse spermatogonial stem cell cryopreservation: feasible protocols and potential biotechnological applications |
| title_short |
Horse spermatogonial stem cell cryopreservation: feasible protocols and potential biotechnological applications |
| title_full |
Horse spermatogonial stem cell cryopreservation: feasible protocols and potential biotechnological applications |
| title_fullStr |
Horse spermatogonial stem cell cryopreservation: feasible protocols and potential biotechnological applications |
| title_full_unstemmed |
Horse spermatogonial stem cell cryopreservation: feasible protocols and potential biotechnological applications |
| title_sort |
horse spermatogonial stem cell cryopreservation: feasible protocols and potential biotechnological applications |
| publisher |
Cell and Tissue Research |
| publishDate |
2020 |
| url |
https://repositorio.inpa.gov.br/handle/1/17005 |
| _version_ |
1787141626643087360 |
| score |
11.653393 |