Sperm separation liquid and preparation method thereof

A technology for separating liquid and sperm, applied in the field of sperm separation, can solve the problems of sperm oxidative damage, affecting the outcome of assisted reproduction, affecting sperm motility and function, etc., to avoid oxidative damage, ensure vitality and DNA integrity.

Inactive Publication Date: 2020-09-01
太东(镇江)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The separation operation process and separation reagents will cause oxidative damage to the sperm, which affects the motility and function of the sperm, and then affects the outcome of subsequent assisted reproduction
Sperm density gradient separation is a commonly used in vitro technique for assisted reproduction. Density gradient separation can effectively remove white blood cells, epithelial cells, dead sperm, deformed or immature sperm and cell debris in semen, but it will also cause oxidative damage to sperm

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Weigh 55.52g NaCl, 3.28g KCl, 0.47g MgSO 4 ·7H 2 O and 2.79 g CaCl 2 2H 2 O, 10.08g NaHCO 3 , 0.46g KH 2 PO 4 , 0.33g C 3 h 3 o 3 Na, 0.72 g C 6 h 12 o 6 , 46.47g HEPES, dissolved in 1.5L water for injection and filtered through a 0.22μm filter; add 0.15g glutathione, 0.9g vitamin C, 0.9g vitamin E, 0.9g β-carotene and 0.9g Add 17.93g C in the gained mixed solution after the catechol is stirred evenly 3 h 5 o 3 Na, stir well and dilute to 2L with water for injection; divide the above solution into two parts, add 4L silane-wrapped colloidal silicon dioxide solution to one part, dilute to 5L, and prepare 80% v / v sperm Separation solution; add 2L silane-coated colloidal silicon dioxide solution to another part, add water for injection to make up to 5L, and configure it as 40% v / v sperm separation solution.

Embodiment 2

[0024] Weigh 58.44g NaCl, 3.43g KCl, 0.49g MgSO 4 ·7H 2 O and 2.94 g CaCl 2 2H 2 O, 10.92g NaHCO 3 , 0.48g KH 2 PO 4 , 0.35g C 3 h 3 o 3 Na, 0.9 g C 6 h 12 o 6 , 47.66g HEPES, dissolved in 1.5L water for injection and filtered through a 0.22μm filter; add 0.31g glutathione, 1g vitamin C, 1g vitamin E, 1gβ-carotene and 1g catechol to the above mixture and stir After uniformity, add 19.05g C in the resulting mixture 3 h 5 o 3 Na, 0.5g gentamycin, 50g human serum albumin, after stirring evenly, dilute to 2L with water for injection; Divide the above-mentioned obtained solution into two parts, add 4L colloidal silicon dioxide solution wrapped in silane to one part, and dilute to 2L with water for injection. The volume is up to 5L, and it is configured as 80% v / v sperm separation solution; another part is added with 2 L of silane-coated colloidal silicon dioxide solution, and water for injection is added to make the volume to 5 L, and it is configured as 40% v / v sperm...

Embodiment 3

[0026] Weigh 61.36g NaCl, 3.58g KCl, 0.52g MgSO 4 ·7H 2 O and 3.09 g CaCl 2 2H 2 O, 11.76g NaHCO 3 , 0.52g KH 2 PO 4 , 0.37g C 3 h 3 o 3 Na, 1.08g C 6 h 12 o 6 , 48.85g HEPES, dissolved in 1.5L water for injection and filtered through a 0.22μm filter; add 0.46g glutathione, 1.1.g vitamin C, 1.1g vitamin E, 1.1gβ-carotene and 1.1 Add 20.17g C in the gained mixed solution after the g catechol is stirred evenly3 h 5 o 3 After the Na is stirred evenly, dilute it to 2L with water for injection; divide the above solution into two equal parts, add 4L colloidal silicon dioxide solution wrapped in silane to one part, dilute to 5L, and configure it for 80% v / v sperm separation solution; another part was added 2L silane-coated colloidal silicon dioxide solution, added water for injection to make up to 5L, and configured as 40% v / v sperm separation solution.

[0027] The separation solution obtained in the above examples was filtered through a 0.22 μm filter membrane again, ...

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PUM

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Abstract

The invention provides a sperm separating liquid, which is characterized by comprising the following components: 13+ / -1 mM NaHCO3, 0.36+ / -0.02 mM KH2PO4, 0.32+ / -0.02 mM C3H3O3Na, 17+ / -1 mM C3H5O3Na, 0.5+ / -0.1 mM C6H12O6, 20+ / -0.5 mM HEPES, 0.1+ / -0.05 mM of glutathione; 0.51+ / -0.06 mM of vitamin C; 0.21+ / -0.02 mM of vitamin E; 0.07+ / -0.02 mM [beta]-carotene; 0.82+ / -0.09 mM of catechol; 100+ / -5mM NaCl, 4.6+ / -0.2 mM KCl, 0.2+ / -0.01 mM of MgSO4.7H2O, 2.0+ / -0.1 mM of CaCl2.2H2O, 5 mg / L of gentamicin, 5 g / L of human serum albumin and a colloidal silicon dioxide solution wrapped by silane with a volume ratio of 80% or 40% and dissolved with water for injection. Glutathione, vitamin C, vitamin E and other antioxidants are added into the sperm separation liquid, so that oxidative stress reaction ofsperms caused by oxidation substances in the sperms and the separation liquid can be avoided, and the vitality of sperms is ensured.

Description

technical field [0001] The invention relates to the field of sperm separation, in particular to a sperm separation liquid and a preparation method thereof. Background technique [0002] The most common cause of infertility in men with normal semen indicators is abnormal sperm DNA, and fertilization ability depends not only on the absolute number of sperm, but also on the function of sperm. Sperm motility is a reliable indicator of sperm fertilization ability, and in assisted reproduction (ART), the number of separable motile sperm in semen directly affects infertility treatment options. The separation operation process and separation reagents will cause sperm to produce oxidative damage, which affects the motility and function of sperm, and then affects the outcome of subsequent assisted reproduction. Sperm density gradient separation is a commonly used in vitro technique for assisted reproduction. Density gradient separation can effectively remove white blood cells, epithe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/076
CPCC12N5/061C12N2500/38C12N2501/999C12N2509/00
Inventor 刘年
Owner 太东(镇江)生物科技有限公司
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