Please use this identifier to cite or link to this item: https://repositorio.inpa.gov.br/handle/1/17005
Title: Horse spermatogonial stem cell cryopreservation: feasible protocols and potential biotechnological applications
Authors: 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
Keywords: 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
Issue Date: 2017
metadata.dc.publisher.journal: Cell and Tissue Research
metadata.dc.relation.ispartof: Volume 370, Número 3, Pags. 489-500
Abstract: 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.
metadata.dc.identifier.doi: 10.1007/s00441-017-2673-1
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