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|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|
Glial Cell Line Derived Neurotrophic Factor Receptor
Glial Cell Line Derived Neurotrophic Factor Receptor Alpha 1
Nanos C2hc Type Zinc Finger 2 Protein
Zinc Finger Protein
Fast Freezing Method
In Vitro Study
Slow Freezing Method
Adult Stem Cell
Adult Germline Stem Cells
|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.|
|Appears in Collections:||Artigos|
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