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Downstream Processing01:29

Downstream Processing

Downstream processing begins once fermentation is complete and involves a series of steps to recover and purify products such as acids, vitamins, antibiotics, or proteins.Cell HarvestingFor example, for intracellular protein-based products, the first step is harvesting the cells. This is typically achieved using centrifugation or filtration to separate the cells from the liquid phase.Cell Disruption for Intracellular ProductsIf the target product is intracellular, the harvested cells must be...

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Antioxidant components of ram seminal plasma are influenced by age, season, and the RsaI polymorphism of the MTNR1A gene.

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Manipulation and In Vitro Maturation of Xenopus laevis Oocytes, Followed by Intracytoplasmic Sperm Injection, to Study Embryonic Development
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Dextran/Swim-Up Procedure for Ram Sperm Selection.

Rosaura Pérez-Pe1, Teresa Muiño-Blanco2, José Álvaro Cebrián-Pérez2

  • 1Grupo BIOFITER-Instituto Universitario de Investigación en Ciencias Ambientales de Aragón (IUCA), Departamento de Bioquímica y Biología Molecular y Celular, Facultad de Veterinaria, Universidad de Zaragoza, Zaragoza, Spain. rosperez@unizar.es.

Methods in Molecular Biology (Clifton, N.J.)
|April 9, 2025
PubMed
Summary

This study introduces a novel dextran/swim-up technique for sperm separation, avoiding centrifugation. This method effectively recovers highly motile and viable ram spermatozoa, crucial for reproductive technologies.

Keywords:
SelectionSeminal plasmaSeparationSpermatozoa

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Area of Science:

  • Reproductive Biology
  • Spermatozoa Separation
  • Animal Reproduction

Background:

  • Spermatozoa separation from seminal plasma is vital for reproductive technologies.
  • Conventional methods often involve centrifugation, which can harm spermatozoa, particularly in rams.
  • Damage to spermatozoa affects their motility and viability, impacting reproductive success.

Purpose of the Study:

  • To develop a non-centrifugation-based technique for separating ram spermatozoa.
  • To achieve high recovery of motile and viable spermatozoa free from seminal plasma.
  • To provide a detailed protocol for a simple, rapid, and high-yield sperm separation method.

Main Methods:

  • A specialized dextran/swim-up technique was developed, eliminating the need for centrifugation.
  • Detailed protocols for media preparation and modified procedures for specific applications are described.
  • The method requires minimal equipment and is adaptable for various mammalian species.

Main Results:

  • The dextran/swim-up technique successfully recovered highly motile and viable ram spermatozoa.
  • Spermatozoa obtained were free from seminal plasma, indicating effective separation.
  • The procedure proved to be simple, rapid, and high-yield.

Conclusions:

  • The developed dextran/swim-up technique offers a superior alternative to centrifugation for sperm separation.
  • This method preserves sperm motility and viability, essential for assisted reproductive technologies.
  • The technique is broadly applicable to spermatozoa from various mammalian species sensitive to centrifugation.