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Isolation and cryopreservation of Pseudopimelodus mangurus (Siluriformes) spermatogonial cells.
Pessoa, Giselle Pessanha; López, Lucia Suárez; Rosero, Jenyffer Mairely; Dos Santos, Silvio Carlos Alves; Yasui, George Shigueki; Senhorini, José Augusto; Monzani, Paulo Sérgio.
Afiliación
  • Pessoa GP; Institute of Bioscience, São Paulo State University, Botucatu, SP, Brazil; Laboratory of Fish Biotechnology, Chico Mendes Institute of Biodiversity Conservation, National Center for Research and Conservation of Continental Aquatic Biodiversity, Pirassununga, SP, Brazil. Electronic address: giselle.p
  • López LS; Institute of Bioscience, São Paulo State University, Botucatu, SP, Brazil; Laboratory of Fish Biotechnology, Chico Mendes Institute of Biodiversity Conservation, National Center for Research and Conservation of Continental Aquatic Biodiversity, Pirassununga, SP, Brazil.
  • Rosero JM; Laboratory of Fish Biotechnology, Chico Mendes Institute of Biodiversity Conservation, National Center for Research and Conservation of Continental Aquatic Biodiversity, Pirassununga, SP, Brazil; Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São P
  • Dos Santos SCA; AES Tiête, Promissão, SP, Brazil.
  • Yasui GS; Laboratory of Fish Biotechnology, Chico Mendes Institute of Biodiversity Conservation, National Center for Research and Conservation of Continental Aquatic Biodiversity, Pirassununga, SP, Brazil; Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of São P
  • Senhorini JA; Institute of Bioscience, São Paulo State University, Botucatu, SP, Brazil; Laboratory of Fish Biotechnology, Chico Mendes Institute of Biodiversity Conservation, National Center for Research and Conservation of Continental Aquatic Biodiversity, Pirassununga, SP, Brazil.
  • Monzani PS; Institute of Bioscience, São Paulo State University, Botucatu, SP, Brazil; Laboratory of Fish Biotechnology, Chico Mendes Institute of Biodiversity Conservation, National Center for Research and Conservation of Continental Aquatic Biodiversity, Pirassununga, SP, Brazil.
Cryobiology ; 116: 104941, 2024 Sep.
Article en En | MEDLINE | ID: mdl-39029551
ABSTRACT
Spermatogonia cryopreservation can be a strategy for future conservation actions. The neotropical Siluriformes Pseudopimelodus mangurus was already classified as vulnerable on the Red List of Threatened Species. P. mangurus spermatogonial cells were isolated, assessed, and cryopreserved. Fragments of the testis were enzymatically dissociated, purified using Percoll density gradient, and submitted to differential plating. Fractionated cells were evaluated by microscopy, ddx4 (vasa) relative expression, and alkaline phosphatase activity. Cryopreservation was conducted using ethylene glycol, glycerol, dimethyl sulfoxide (DMSO), dimethylacetamide (DMA), and propanediol at 1 M, 1.5 M, and 2 M. Cell viability was evaluated and cell concentration was determined. Cell fractions from 20 % and 30 % Percoll gradient bands showed the highest concentrations of spermatogonia. The fraction mix showed 54 % purity and 93 % viability. After differential plating, 60 % purity and 92 % viability were obtained. Spermatogonial cells showed high alkaline phosphatase activity compared to spermatocytes and spermatids. The relative spermatogonial ddx4 expression from the Percoll density gradient was about twice as high as in samples from the testis and the differential plating. The increased ddx4 expression indicated the enrichment of spermatogonial cells by density gradient step and dead cells expressing ddx4 in differential plating, or ddx4 decreasing expression during cell culture. For this reason, cells from the Percoll gradient were chosen for cryopreservation. Propanediol at 1 M demonstrated the best condition for spermatogonial cell cryopreservation, presenting 98 % viability, while dimethylacetamide at 2 M represented the least favorable condition, with approximately 47 % viability. These findings are essential for P. mangurus spermatogonial cell cryopreservation, aiming to generate a spermatogonia cryobank for future conservation efforts.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espermatogonias / Bagres / Criopreservación / Supervivencia Celular Límite: Animals Idioma: En Revista: Cryobiology Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espermatogonias / Bagres / Criopreservación / Supervivencia Celular Límite: Animals Idioma: En Revista: Cryobiology Año: 2024 Tipo del documento: Article Pais de publicación: Países Bajos