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Methods and systems for reducing dna fragmentation in a processed sperm sample

A sample, sperm technology, applied in the field of reducing DNA fragmentation in processed sperm samples and systems to address issues such as damage

Inactive Publication Date: 2012-12-05
INGURAN LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, since spermatozoa are often fragile cells, sperm samples obtained after withdrawal from ejaculation may suffer additional iatrogenic damage during most sperm processing (while preparing samples for insemination)

Method used

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  • Methods and systems for reducing dna fragmentation in a processed sperm sample
  • Methods and systems for reducing dna fragmentation in a processed sperm sample
  • Methods and systems for reducing dna fragmentation in a processed sperm sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 - The first experiment was performed to analyze the difference in the amount of DNA fragmentation before and after sex sorting. Sperm samples were taken from 5 Jersey bulls and each sample was divided into two aliquots. The first set of aliquots were sex sorted and then frozen. A second set of aliquots was directly frozen. Table 1 shows the level of DNA fragmentation obtained for each bull before and after sorting.

[0068] Table 1 (% DNA fragmentation, sex sorting)

[0069] refer to

[0070]Baseline levels of DNA damage in pre-sort samples from the 5 bulls ranged from 5.3% to 11%, with a mean and standard deviation of 7.9±2.1. The level of sperm DNA fragmentation obtained in sex-sorted sperm samples was much lower with a mean and standard deviation of 3.1 ± 1.9. The average reduction in sperm DNA fragmentation was 63%, but the reduction in Bull 2 ​​was as high as 85%.

Embodiment 2

[0071] Example 2 - The second experiment looks specifically at DNA fragmentation in each sorted subpopulation after sex sorting. Five more jersey bulls were used in this experiment; each was collected and sorted to obtain three sperm subpopulations. The first sperm subpopulation consisted mainly of those sperm considered dead, indicated by red food dye or propidium iodide via conventional sorting techniques. The second and third subpopulations consisted mainly of live sorted spermatids. A portion of each sample was tested prior to sorting to establish a baseline for DNA fragmentation. As shown in Table 2, the baseline of DNA fragmentation had a mean and standard deviation of 7.9 ± 2.5. Averaged across bulls, DNA fragmentation assays in the sorted X-chromosome-bearing subpopulation had a mean and standard deviation of 1.8±1.5 and a mean and standard deviation of 1.2±0.6 in the Y-chromosome-bearing subpopulation. The third sperm subpopulation contained all dead sperm, which...

Embodiment 3

[0074] Example 3 - A third experiment was performed to analyze the distribution of sperm DNA fragmentation in 100 sex-sorted sperm storage tubes after thawing to compare the variation between samples collected at different times from 10 Holstein bulls. Each sperm storage tube was collected and sex-sorted for X-chromosome-carrying sperm. As can be seen from Table 3, sperm storage tubes collected from the same bull on different days often have very similar DNA breaks. Although there were some occasional outliers, most samples obtained from individual bulls showed similar DNA fragmentation regardless of whether they were collected on different days.

[0075] Table 3 (% DNA fragmentation, X-sorted samples collected on different days)

[0076]

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PUM

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Abstract

A method and system for processing reproductive cell samples and for sorting sperm with reduced levels and occurrences of DNA fragmentation compared to conventional sorting and processing methods, and for using reproductive cell and sperm cell samples with low levels of DNA fragmentation to improve viability of insemination samples fertility and success rates of assisted reproductive procedures, including artificial insemination, in vitro fertilization, intracytoplasmic injection, and other related techniques.

Description

[0001] This international PCT (Patent Cooperation Treaty) application is a continuation-in-part of, and claims priority from, U.S. Provisional Patent Application 61 / 320,183 filed April 1, 2010, U.S. Provisional Patent Application Patent Application 61 / 324,192 and International PCT Application No. PCT / US10 / 54549, each of which is hereby incorporated by reference. technical field [0002] Embodiments of the invention generally relate to methods and systems for reducing the number of DNA fragmentation events in various populations of treated cells and sperm cells, and more specifically, to methods for modifying cell processing and sperm sorting processes to reduce DNA fragmentation events in various cell and sperm suspensions, methods and systems for reducing the incidence of DNA fragmentation in various cell and sperm samples, and for improving the overall success of assisted reproductive techniques and procedures. Background technique [0003] Sperm and sex-sorted sperm (sper...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/076C12N5/02C12Q1/02G01N33/48
CPCG01N33/5091
Inventor J·莫雷诺M·埃文斯M·杰兰德J·格萨维兹C·冈萨雷斯·马丁C·洛佩兹·费尔南德斯
Owner INGURAN LLC
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