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Sperm suspensions for sorting into x or y chromosome-bearing enriched populations

A suspension and sperm technology, applied in the preservation of human or animal bodies, the preparation and application of test samples, etc., can solve the problems of reduced forward motility

Active Publication Date: 2007-08-08
INGURAN LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Every step of the staining and sorting process is stressful for sperm cells, resulting in reduced sperm cell viability or motility, especially forward motility

Method used

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  • Sperm suspensions for sorting into x or y chromosome-bearing enriched populations
  • Sperm suspensions for sorting into x or y chromosome-bearing enriched populations
  • Sperm suspensions for sorting into x or y chromosome-bearing enriched populations

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] Bull semen was collected from sexually mature bulls using artificial vaginas, and samples were transferred to the staining facility in temperature-controlled containers at 25°C. Once received, the semen was analyzed for concentration, motility using the Hamilton-Thorn Motility Analyzer (IVOS) according to standard and well-known methods (Farrell et al. Theriogenology, 49(4):871-9 (March 1998)). and forward mobility. Depending on the semen concentration, suspend the semen in TCA buffer or carbonate-based inhibitors and prepare several tubes containing 150 × 10 6 Sperm / ml of suspension. Table I below shows the compositions and staining conditions used.

[0101] Table I

[0102] Sample name

[0103] An aliquot of 10 mM Hoechst in water was added to the sperm suspension to give a concentration of 600 µM Hoechst. The sperm suspension was left in a 28°C water bath during staining (approximately 1 hour). A 50 μl aliquot was pipetted from the stained sperm suspen...

Embodiment 2

[0105]Bull semen was collected from sexually mature bulls using artificial vaginas, and samples were transferred to the staining facility in temperature-controlled containers at 25°C. Once received, the semen was analyzed for concentration, motility using the Hamilton-Thorn Motility Analyzer (IVOS) according to standard and well-known methods (Farrell et al. Theriogenology, 49(4):871-9 (March 1998)). and forward mobility. Depending on the semen concentration, suspend the semen in TCA buffer or carbonate-based inhibitors and prepare several tubes containing 450 × 10 6 Sperm / ml of suspension. Table II below shows the compositions and staining conditions used.

[0106] Table II

[0107] Sample name

[0108] An aliquot of 10 mM Hoechst in water was added to the sperm suspension to give a concentration of 1000 µM Hoechst. The sperm suspension was left in a water bath at 28°C for 1 hour, and then diluted to 150×10 with TCA containing 10 mM pyruvate or a carbonate-based...

Embodiment 3

[0110] Bull semen was collected from sexually mature bulls using artificial vaginas, and samples were transferred to the staining facility in temperature-controlled containers at 25°C. Once received, the semen was analyzed for concentration, motility using the Hamilton-Thorn Motility Analyzer (IVOS) according to standard and well-known methods (Farrell et al. Theriogenology, 49(4):871-9 (March 1998)). and forward mobility. Depending on the semen concentration, suspend the semen in TCA buffer or carbonate-based inhibitors and prepare several tubes containing 450 × 10 6 Sperm / ml of suspension. Table II below shows the compositions and staining conditions used.

[0111] Table III

[0112] Sample name

[0113] An aliquot of 10 mM Hoechst in water was added to the sperm suspension to give a concentration of 300 µM Hoechst. The sperm suspension was left in a water bath at 41°C for 30 minutes and then diluted to 150×10 with TCA containing 10 mM pyruvate or a carbonate-b...

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Abstract

Sperm cell suspensions comprising a motility inhibitor are disclosed. The cells contained in such suspensions tend to have a greater capacity for enduring the various process steps typically associated with the sorting of sperm cells into gender enriched populations, thereby resulting in post-sort compositions with an increased number of viable or motile sperm. Processes for forming such cell suspensions, as well as processes for staining sperm cells, are also disclosed.

Description

field of invention [0001] The present invention generally relates to methods for sorting sperm cells. More specifically, the present invention relates to the preparation of a suspension of sperm cells with reduced motility relative to ejaculated sperm, more specifically temporarily reduced motility; said suspension may be used, for example, to sort sperm cells bearing X or Method for Enriching Sperm Cell Populations for the Y Chromosome. Background of the invention [0002] Fertilization of animals by artificial insemination (AI) and embryo transfer after in vitro fertilization is a well-established technology. In the livestock industry, there are clear advantages in being able to influence reproductive outcome so that offspring possess one or more desired characteristics. For example, in the dairy industry, if the offspring can be pre-selected in favor of females and the production of cows can be guaranteed, it will bring economic benefits. Separating sperm into enriched...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06A61K35/52G01N1/30A01N1/02
Inventor C·L·路德维格K·S·克劳利C·N·格拉维斯
Owner INGURAN LLC
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