Method for improving sperm motility with Sertoli cells as trophoblast

A technology of Sertoli cells and sperm motility, applied in the field of sperm in vitro treatment, can solve the problem of difficult to judge whether it is live sperm, sperm can not move, etc., and achieve the effect of increasing fertilization ability, vitality and viability.

Inactive Publication Date: 2017-05-24
QINGDAO AGRI UNIV
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Problems solved by technology

However, the sperm in the testis cannot move, and it is difficult to judge whether it is a live

Method used

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  • Method for improving sperm motility with Sertoli cells as trophoblast
  • Method for improving sperm motility with Sertoli cells as trophoblast
  • Method for improving sperm motility with Sertoli cells as trophoblast

Examples

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Embodiment 1

[0028] This embodiment is an experimental group, and the processing of the experimental group is as follows:

[0029] A method for improving sperm motility with Sertoli cells as a trophoblast, comprising the following steps:

[0030] (1) Take out the testes of mice 4-6 days after birth, wash them in normal saline with a preheated temperature of 37°C, remove excess fat in PBS containing double antibodies, and peel off the white membrane. The testis was transferred to DMEM / F12, mixed by pipetting and centrifuged to remove the supernatant;

[0031] (2) Add 1.5ml hyaluronidase and 1.5ml collagenase IV to digest for 2 minutes, add an equal volume (3ml) of DMEM / F12, blow and mix, and centrifuge to remove the supernatant;

[0032] (3) Add 1.5ml of trypsin and 1.5ml of DNase to digest for 2 minutes, add an equal volume (3ml) of DMEM / F12 culture medium containing 10% FBS to stop the digestion, and pass the cell suspension through 80 mesh, 200 mesh, and 300 mesh successively Sieve the...

Embodiment 2

[0038] This embodiment is the specific implementation of the control group, and the processing of the control group is as follows:

[0039] (1) Mature male mice were killed by dislocation, and the testes and epididymis were removed immediately, and the blood and adipose tissue were removed on sterile filter paper;

[0040] (2) Cut the cauda epididymis with ophthalmic scissors, take out the sperm mass, add 1ml of DMEM / F12 culture system containing 10% FBS, place in an incubator at 37°C, and incubate for 2 hours.

[0041] (3) After culturing for 2 hours, use an automatic sperm analyzer to record the motility of the sperm.

[0042] Such as figure 1 and figure 2 Shown is the picture of the results of the automatic sperm analyzer, the red line represents fast forward motility sperm, the green line represents slow forward motility sperm, the blue line represents non-progressive motility sperm, and the yellow dot represents immobile sperm.

[0043] Such as image 3 and Figure ...

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Abstract

The invention relates to a method for improving the sperm motility with Sertoli cells as the trophoblast. The method mainly includes the steps of: (1) conducting isolation culture on Sertoli cells; (2) preparing a Sertoli cell trophoblast: conducting isolation culture on Sertoli cells for 2 days to form a cell monolayer, adding mitomycin C to perform treatment for 2h so as to prepare a Sertoli cell trophoblast; and (3) acquiring sperm from an epididymis of a mature male animal, adding the sperm into a culture system adopting Sertoli cells as the trophoblast, and performing co-culture on the sperm and Sertoli cells. The method provided by the invention adopts Sertoli cells as the trophoblast for in-vitro culture of sperms in testicle and epididymis, solves the problem of low in-vitro sperm motility, enhances the motility and survival rate of in-vitro sperms, and increases the fertilization capability of sperms.

Description

technical field [0001] The technical invention belongs to the technical field of extracorporeal treatment of sperm, and specifically relates to a method for improving sperm motility by using Sertoli cells as a trophoblast. Background technique [0002] In vitro fertilization refers to the technology in which mammalian sperm and eggs complete the fertilization process in an artificially controlled environment outside the body. In biology, animals obtained by transplanting in vitro fertilized embryos into mothers are called test-tube animals. Artificial insemination technology is a high-tech applied to the breeding of mammals. Using artificial insemination combined with embryo transfer technology, it can better develop the reproductive potential of female animals with excellent genetic characteristics and quickly expand the herd of fine breeds. [0003] Since the birth of the first test-tube baby in 1978, breakthroughs have been made in the treatment of infertility and infert...

Claims

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Application Information

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IPC IPC(8): C12N5/076
CPCC12N5/061C12N2502/04
Inventor 董焕声杜淼王明明李桢潘庆杰
Owner QINGDAO AGRI UNIV
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