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Related Experiment Videos

A simple method for preparing modified TES and Tris yolk buffer that is optically clear and membrane filterable

P S Weathersbee1, A R Johnson

  • 1Irvine Scientific, Santa Ana, California.

Fertility and Sterility
|April 1, 1995
PubMed
Summary

A new modified TES and Tris (TEST) yolk buffer (TYB) using egg yolk extract performed identically to the traditional whole egg yolk preparation in sperm function tests. This lecithin-rich extract offers an effective alternative for laboratory use.

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Area of Science:

  • Reproductive biology
  • Sperm function analysis
  • Biochemical analysis of media

Background:

  • Traditional TES and Tris (TEST) yolk buffer (TYB) is crucial for sperm preparation in assisted reproduction.
  • The preparation of TYB traditionally uses whole egg yolk, posing potential variability and challenges.
  • Evaluating alternative, standardized components for TYB is essential for consistent laboratory performance.

Purpose of the Study:

  • To assess the laboratory performance of a modified TYB using a commercial egg yolk extract.
  • To compare the efficacy of modified TYB with the traditional whole egg yolk TYB.
  • To characterize the phospholipid content, specifically lecithin, of the modified TYB formulation.

Main Methods:

  • Sperm samples from normozoospermic and oligozoospermic males were stored in modified or commercial TYB.

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  • Sperm function was evaluated using the optimized sperm penetration assay (SPA).
  • Sperm swim-up recovery rates and average ovum penetration were measured; HPLC analyzed egg yolk extract composition.

    • Main Results:

    • No significant differences were observed in sperm swim-up recovery rates between modified and commercial TYB.
    • Optimized sperm penetration assay (SPA) results were comparable for both TYB preparations across different semen qualities.
    • High-pressure liquid chromatography (HPLC) identified lecithin as the primary component of the commercial egg yolk extract.

    Conclusions:

    • Commercially available egg yolk extract is a rich source of lecithin and can be filtered (0.2-microns).
    • Modified TYB prepared with egg yolk extract demonstrates equivalent performance to whole egg yolk TYB.
    • The commercial egg yolk extract serves as an effective and potentially more standardized substitute for whole egg yolk in TYB preparation.