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In vitro human sperm culture solution for improving sperm motility and application thereof

A technology for motility and in vitro cultivation, applied in the field of culture medium, can solve the problems of insufficient improvement of sperm motility and low success rate of artificial insemination, and achieve the effects of improving low sperm motility, obvious improvement of vitality, and avoiding toxic effects

Active Publication Date: 2013-10-16
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But the obvious disadvantage is that the improvement of sperm motility is not enough, the success rate of artificial insemination has not been high (about 10%) may also be related to this

Method used

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  • In vitro human sperm culture solution for improving sperm motility and application thereof

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Embodiment 1

[0024] 1. Sample Preparation

[0025] Sperm samples from patients with severe asthenozoospermia were selected for the experiment. Semen ejaculation by masturbation, using a specific sterile container to collect sperm samples after 2-7 days of abstinence. The sample water bath was kept at 22-37°C, and the semen after liquefaction was evaluated according to the WHO1999 evaluation standard, using the computer-assisted semen analyzer CASA (IVOS; Hamilton-Thorn Research, Inc. , sperm count, percentage of motility and motility) were assessed. Asthenozoospermia is defined as grade A sperm count <25% or grade A+B sperm count <50%, (World Health Organization, 1999), while all other parameters are normal. Specimens containing white blood cells or somatic cell contamination were excluded based on morphological observations. The motility parameters of severe asthenozoospermia sperm are (a+b) grade <25%, of which a grade sperm <5%.

[0026] 2. In vitro sperm culture

[0027] 1) Densit...

Embodiment 2

[0039] 1. Sample Preparation

[0040] Sperm samples from patients with moderate to mild asthenozoospermia were selected for the experiment. Semen ejaculation by masturbation, using a specific sterile container to collect sperm samples after 2-7 days of abstinence. The sample water bath was kept at 22-37°C, and the liquefied semen was evaluated according to the WHO1999 evaluation standard, using the computer-assisted semen analyzer CASA (IVOS; Hamilton-Thorn Research, Inc. Beverly, MA, USA), and the sperm parameters (volume , sperm count, percentage of motility and motility) were assessed. According to the World Health Organization standard in 1999, the sperm motility parameters of moderate and mild asthenozoospermia are 25%<(a+b) grade <50% and a grade sperm <25%.

[0041] 2. In vitro sperm culture

[0042] 1) Density gradient centrifugation separates sperm from semen.

[0043] Place 1 mL of 60% (v / v) Percoll separation solution in the test tube, layer 1 mL-1.5 mL of semen...

Embodiment 3

[0051] 1. Sample Preparation

[0052] Sperm samples from healthy male sperm donors are selected for the experiment. Semen ejaculation by masturbation, using a specific sterile container to collect sperm samples after 2-7 days of abstinence. The sample water bath was kept at 22-37°C, and the semen after liquefaction was evaluated according to the WHO1999 evaluation standard, using the computer-assisted semen analyzer CASA (IVOS; Hamilton-Thorn Research, Inc. Beverly, MA, USA), and the sperm parameters (volume , sperm count, percentage of motility and motility) were assessed.

[0053] 2. In vitro sperm culture

[0054] 1) Density gradient centrifugation to sort sperm from semen.

[0055] Place 1mL of 60% (v / v) Percoll separation solution in the test tube, slowly spread 1mL-1.5mL of semen on it, centrifuge at 300g for 5-10 minutes, and discard the supernatant. Resuspend the sperm suspension in 2mL of HTF-S culture medium, centrifuge at 300g for 5-10 minutes, repeat this step ...

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Abstract

The invention discloses in vitro human sperm culture solution for improving sperm motility and application thereof. The culture solution contains fructose 6-phosphoric acid (F6P). The in vitro human sperm culture solution for improving the sperm motility of human sperms is used for culturing the purified human sperms which leave from a human body; the vitality of normal human sperms is capable of maintaining sperm movement and can be obviously improved; and a meaningful reference is provided for improving and assisting the reproductive technology. The F6P is directly used for cultivating and diluting the culture solution, and does not relate to organic solvents such as ethanol or dimethylsulfoxide (DMSO) and the like; the toxic action of the ethanol or the DMSO on the sperms is avoided; by adopting the F6P added to the sperm culture solution, the vitality of the sperms of clinical patients with asthenospermia can be obviously improved; the improvement degree of the vitality is the most obvious for severe oligospermia; the activity of a part of cultured sperms reaches the normal sperm vitality range; and the improvement degree of the vitality for the severe oligospermia is the most obvious. This fully demonstrates that low motility of the sperms of the patients with asthenospermia can be improved by using of the F6P.

Description

technical field [0001] The invention relates to a culture solution, in particular to a culture solution for improving the motility of in vitro cultured sperm and its application. [0002] Background technique [0003] At present, the infertility rate accounts for about one tenth of couples of reproductive age, of which male factor infertility accounts for about half, but more than 85% of infertile men can produce sperm. An important clinical manifestation of male infertility is asthenozoospermia, that is, low sperm motility, leading to male infertility. Improving sperm motility is an important method to treat this type of male infertility. This also shows the importance of sperm motility in the normal function of sperm. [0004] There are a variety of assisted reproductive methods for the treatment of infertility, such as artificial insemination (IUI), in vitro fertilization-embryo transfer (IVF), and intracytoplasmic sperm injection (ICSI), all of which involve the cultiv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/076
Inventor 沙家豪周作民郭雪江牛鑫
Owner NANJING MEDICAL UNIV
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