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Coating carrier and its use in method for sperm maturity detection and non-invasive mature sperm separation

A carrier and coating technology, applied in biochemical equipment and methods, bioreactors/fermenters for specific purposes, analytical materials, etc., can solve problems such as embryo stop development, embryo instability, deformity, etc., and achieve high maturity , The results are accurate and reliable, and the effect of ensuring safety

Inactive Publication Date: 2014-06-04
SHENZHEN PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Assisted reproduction is an effective method to treat infertility, but at the same time there are certain genetic risks, especially when using intracytoplasmic sperm injection (ICSI) technology, the sperm that penetrate into the egg cell is artificially selected, and the selection has a certain degree of randomness. The maturity of the selected sperm will determine the health of the embryo. After the immature sperm combines with the egg cell, the embryo will become unstable, which will easily lead to serious consequences such as miscarriage, embryo stop development, and deformity.

Method used

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  • Coating carrier and its use in method for sperm maturity detection and non-invasive mature sperm separation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1: Preparation of coated carrier

[0069] Prepare the coated carrier as follows:

[0070] a. wash the carrier to be coated with ethanol, and dry;

[0071] b. Fully dissolve the obtained hyaluronic acid 2mg / ml and chitosan 8mg / ml with 0.02mol / L citric acid buffer solution (pH 4.0), stir and react at room temperature for 2 hours;

[0072] c. Coated in the carrier;

[0073] d. Wash 3 times with a washing liquid mixed with 10% acetone, 0.5% 1,2,7,8-diepoxyoctane, 25% isopropanol, and the rest of distilled water, and dry.

[0074] Wherein, the coated carrier can be a coated slide, a coated container, a slide with coated spheres or cylinders or cones or cubes or irregular bodies placed or fixed on the surface, and coated or fixed inside. A sphere or cylinder or a cube or an irregular container, etc.

[0075] Wherein, the coating carrier may be a glass carrier, a plastic carrier or a carrier made of other materials.

[0076] Wherein, the coating carrier can be var...

Embodiment 2

[0077] Example 2: Preparation of coated carrier

[0078] The method is the same as in Example 1, except that hyaluronic acid derivatives are used in step b: 6 mg / ml of hyaluronic acid derivatives, 6 mg / ml of chitosan, and 0.10 mol / L of citric acid buffer (pH 4.0 ) fully dissolved, stirred and reacted at room temperature for 2 hours;

[0079] The composition of the washing liquid in step d is: 30% acetone, 3.5% 1,2,7,8-diepoxyoctane, 10% isopropanol, and distilled water as the balance.

[0080] The types of hyaluronic acid derivatives include: the carboxyl group, hydroxyl group, N-acetyl group and reducing end of the hyaluronic acid molecule are formed by chemical modification with other compounds such as esterification, cross-linking, and grafting. derivative compounds.

Embodiment 3

[0081] Example 3: Preparation of coated carrier

[0082] The method is the same as in Example 1, except that, in the step b, hyaluronic acid sodium salt and other buffers are used: hyaluronic acid sodium salt 4mg / ml, chitosan 3mg / ml, 0.06mol / L acetic acid The buffer (pH 4.0) was fully dissolved and mixed, and stirred at room temperature for 2 hours;

[0083] The composition of the washing liquid in the step d is: 20% acetone, 2.0% 1,2,7,8-diepoxyoctane, 15% isopropanol, and distilled water as the balance.

[0084] The above-mentioned sodium hyaluronic acid salt can be replaced by potassium hyaluronic acid salt, and the others are the same.

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Abstract

The invention discloses a coated carrier and the application thereof in the method for detecting maturity of sperms and separating mature sperms noninvasively; the coated carrier is a carrier coated by hyaluronic acid, or hyalurate or hyaluronic acid derivatives; the detecting method is as follows: preparing specimens; loading the specimens to the coated carrier and covering with microscopic glass; reacting for 5-60 minutes at room temperature; observing by a microscope and counting; and calculating the combination ratio of hyaluronic acid of active sperms. The separating method is as follows: preparing specimens; loading the specimens to the coated carrier, reacting for 5-120 minutes at the temperature of 18-37 DEG C; and collecting the combined sperms. The coated carrier has the advantages of precise and reliable results, simple and fast operation, low cost and good safety performance, and can be used for estimating sperms quality, sperms maturity and analysis of the causes of sterility disease of mails, and is applicable to assisting reproduction or reproduction research center to safely collect mature and active sperms, so that the goal of improving embryo quality greatly is achieved.

Description

technical field [0001] The invention relates to the field of detecting sperm quality in vitro and assisted reproduction in vitro, in particular to a coated carrier and its application in methods for detecting sperm maturity and noninvasively separating mature sperm. Background technique [0002] Hyaluronic acid is the main component of the mucous matrix of the egg and zona pellucida. Sperm acrosome and cytoplasmic membrane contain hyaluronidase. When sperm penetrate the cumulus complex, hyaluronidase in the plasma membrane hydrolyzes the mucus hyaluronic acid in the cumulus complex, thereby passing through the mucus; sperm release hyaluronic acid when the acrosome reaction is induced in the zona pellucida The enzyme hydrolyzes hyaluronic acid, which helps sperm penetrate the zona pellucida to complete fertilization. In the whole process, the combination of sperm and hyaluronic acid played an important role. Studies have shown that only mature motile sperm can combine with...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/544C12N5/08C12M1/00
Inventor 刘瑜程锦珍程锦娥
Owner SHENZHEN PEOPLES HOSPITAL
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