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Isolation and cryopreservation of Pseudopimelodus mangurus (Siluriformes) spermatogonial cells

  • 0Institute 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 +

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July 19, 2024

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July 19, 2024

View abstract on PubMed

Summary

This summary is machine-generated.

Cryopreservation of spermatogonia from the vulnerable fish Pseudopimelodus mangurus was optimized using propanediol. This research establishes a method for creating a cryobank to aid future conservation of this species.

Area Of Science

  • Ichthyology
  • Reproductive Biology
  • Conservation Genetics

Background

  • The neotropical fish Pseudopimelodus mangurus is listed as vulnerable.
  • Spermatogonia cryopreservation offers a strategy for species conservation.
  • Effective cryopreservation protocols are needed for P. mangurus.

Purpose Of The Study

  • To isolate, characterize, and cryopreserve spermatogonial cells from P. mangurus.
  • To evaluate different cryoprotective agents and concentrations.
  • To establish optimal conditions for spermatogonial cell cryopreservation for conservation.

Main Methods

  • Testis fragments were dissociated and spermatogonia purified using Percoll density gradients.
  • Cells were assessed via microscopy, alkaline phosphatase activity, and ddx4 gene expression.
  • Cryopreservation trials used ethylene glycol, glycerol, DMSO, DMA, and propanediol at varying molarities.
  • Cell viability and concentration were determined post-thaw.

Main Results

  • Percoll gradient fractions yielded enriched spermatogonial cell populations with high purity and viability.
  • Spermatogonial cells exhibited high alkaline phosphatase activity and ddx4 expression.
  • 1 M propanediol resulted in the highest post-thaw viability (98%).
  • 2 M dimethylacetamide yielded the lowest viability (47%).

Conclusions

  • Percoll gradient purification is effective for isolating viable spermatogonia.
  • Propanediol at 1 M is the optimal cryoprotective agent for P. mangurus spermatogonia.
  • These findings support the development of a P. mangurus spermatogonia cryobank for conservation.

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