Sperm washing upstream method

Abstract

The invention belongs to the technical field of sperm optimization, and particularly relates to a sperm washing upstream method. The method comprises the following steps: (1) balancing a centrifugal tube A and a centrifugal tube B which are used for containing a fertilization culture solution in a CO2 incubator with the temperature of 37 DEG C and the volume ratio of 5% for 6-18 hours; (2) liquefying the semen at 25-37 DEG C for 30-60 minutes to obtain semen volume, sperm concentration, forward sperm percentage and forward sperm concentration; (3) adding the semen into the centrifugal tube B, and centrifuging; (4) adding part of the fertilization culture solution in the centrifugal tube A into a centrifugal tube B, and centrifuging; (5) adding the residual fertilization culture solution in the centrifugal tube A into the centrifugal tube B, and carrying upstream culture in a CO2 incubator with the temperature of 37 DEG C and the volume ratio of 5% for 20-30 minutes; (6) sucking out upstream liquid of the centrifugal tube B to obtain a sperm suspension after upstream, and obtaining sperm concentration, forward sperm percentage, forward sperm concentration and active sperm percentage; and (7) preserving the precipitate of the centrifugal tube B at room temperature. According to the method, the semen quality is improved, and the problems of low sperm density, poor vitality, low survival rate and the like during in-vitro insemination are solved.

Description

technical field [0001] The invention belongs to the technical field of sperm optimization, and in particular relates to an upstream method of sperm washing. Background technique [0002] Human assisted reproductive technology refers to the use of medical technology and methods to artificially manipulate gametes (sperm and eggs), zygotes (fertilized eggs), and embryos to achieve the purpose of conception. It is divided into artificial insemination and in vitro fertilization-embryo transfer technology and its Various derivative technologies. Sperm washing and optimization is a very important link in the process of human assisted reproductive technology. After in vitro sperm collection, the semen is washed to remove the factors that affect fertilization in the semen, such as white blood cells, viruses, bacteria, and sperm in the semen. Motility impurities, etc., so as to obtain high-quality, high-viability sperm, and high-viability sperm can improve the success rate of sperm a...

AI Technical Summary

Problems solved by technology

[0003] The object of the present invention is to provide a sperm washing upstream method, which can separate high motility sperm from dead sperm, poor motility sperm, agglutinated sperm, deformed sperm Or other harmful components, impurities, etc., so as to screen out the sperm with good shape, high vitality, and high quality, so that the sperm with high vitality can be redistributed, improve the quality of semen, and solve the problem of low sperm density, poor vitality, and survival during in vitro fertilization. Low rate and other issues