Culture solution for bovine in vitro fertilization (IVF) and method for improving bovine IVF

A technology of culture fluid and bovine body, applied in biochemical equipment and methods, culture process, tissue culture, etc., can solve the problems of limited efficiency of in vitro fertilization

Active Publication Date: 2021-02-05
天津力牧生物科技有限公司
View PDF7 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] However, the efficiency of bovine frozen semen for in vitro fertilization with oocytes remains limited

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1: the culture method of bovine in vitro fertilization embryo (ex vivo collection)

[0059] In the present embodiment, in step (1), the collected oocytes were placed in transport medium containing HEPES at 38.8° C. without carbon dioxide supply, and transported back to the laboratory within 24 hours.

[0060] 1. Reagents

[0061] In the specific test of the present invention, if not otherwise specified, the relevant reagents used are described in detail as follows:

[0062] The transportation culture fluid containing HEPES used in this example contains: glycine 50.0mg / L, L-alanine 25.0mg / L, L-arginine hydrochloride 70.0mg / L, L-aspartic acid 30.0mg / L, L-cystine dihydrochloride 26.0mg / L, L-glutamic acid 75.0mg / L, L-glutamine 100.0mg / L, L-histidine hydrochloride monohydrate 21.88mg / L, L-hydroxyproline 10.0mg / L, L-isoleucine 40.0mg / L, L-leucine 60.0mg / L, L-lysine hydrochloride 70.0mg / L, L-methionine 15.0mg / L, L-phenylalanine 25.0mg / L, L-proline 40.0mg / L, L-ser...

Embodiment 1A

[0100] Embodiment 1A: the culture method of bovine in vitro fertilization embryo (ex vivo collection)

[0101] With reference to the method of Example 1, tests were carried out on three kinds of cattle, Holstein (dairy breed), Simmental (beef breed), and Chinese buffalo (serving breed), respectively. The results showed that the cleavage rate was 85-90%. Within the range, the morula rate was in the range of 62-67%, the blastocyst rate was in the range of 50-55%, the apoptosis rate was in the range of 4-6%, and the blastocyst hatching rate on the 9th day was 72-5%. Within the range of 77%; the maturation rates of oocytes in vitro maturation of Holstein, Simmental, and Chinese buffalo were 83.3%, 80.6%, and 75.6%, respectively.

Embodiment 1B

[0102] Embodiment 1B: the culture method of bovine in vitro fertilization embryo (in vitro collection)

[0103] With reference to the method of Example 1, four kinds of cattle were tested respectively for Chinese yellow cattle (Nanyang cattle, service breed), Holstein cattle (dairy breed), Simmental cattle (beef breed), and Chinese water buffalo (service breed), However, the formulations of the fertilization culture medium and the semen preparation culture medium were respectively modified, and the remaining test fluids and operations remained unchanged. The formula of the fertilization medium was changed to: containing 112.0mM sodium chloride, 4.02mM potassium chloride, 2.25mM calcium chloride dihydrate, 0.52mM magnesium chloride hexahydrate, 0.83mM potassium dihydrogen phosphate, 37.0mM sodium bicarbonate, 1.25 mM sodium pyruvate, 10μg / ml heparin, 4mg / ml bovine serum albumin (BSA), 100U / ml penicillin, 100μg / ml streptomycin, 10umoL / L α-lipoic acid, 25μg / ml folic acid, 3mg / ml ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a culture solution for bovine IVF and a method for improving bovine IVF. The method for bovine IVF comprises the following steps: washing a mature cumulus-oocyte complex in afertilization culture solution, transferring the mature cumulus-oocyte complex into the fertilization culture solution, and putting the fertilization culture solution into an incubator for later use;taking a frozen seminal tubule from liquid nitrogen, performing unfreezing in a water bath at 37 DEG C, injecting seminal fluid into a centrifuge tube containing a seminal fluid preparation culture solution, and discarding a supernatant after centrifugation; adding the seminal fluid preparation culture solution into the centrifuge tube, resuspending sperm precipitate, and taking a proper sperm suspension for sperm counting; and adding the sperm suspension with the calculated volume into fertilization culture liquid drops containing oocytes, placing a culture tray into the incubator, and carrying out sperm-ovum co-incubation to complete IVF operation for embryo in-vitro culture. The fertilization culture solution and the seminal fluid preparation culture solution contain sodium pyruvate, heparin and other components. The method can significantly improve the efficiency of bovine IVF.

Description

technical field [0001] The invention belongs to the technical field of animal reproduction, and relates to a technology for agricultural-animal husbandry and veterinary reproduction, in particular to a method for bovine in vitro fertilization. In addition, the present invention also relates to the use of related working test solutions used in bovine in vitro fertilization. Application in fertilization. The bovine in vitro fertilization method of the invention has excellent technical effects. Background technique [0002] In Vitro Fertilization (In Vitro Fertilization) or (external fertilization) refers to the technology in which mammalian sperm and eggs complete the fertilization process in an artificially controlled environment outside the body. It is referred to as IVF in English. Because it is inseparable from embryo transfer technology (ET), it is also referred to as IVF-ET. In biology, animals obtained by transplanting in vitro fertilized embryos into mothers are call...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/075C12N5/076A61D19/02
CPCA61D19/02C12N5/0609C12N5/061C12N2500/12C12N2500/14C12N2500/20C12N2500/24C12N2500/32C12N2500/33C12N2500/34C12N2500/35C12N2500/36C12N2500/38C12N2500/40C12N2501/105C12N2501/11C12N2501/30C12N2501/31C12N2501/39C12N2501/392C12N2501/999
Inventor 王娜王小武郝少强赵明礼韩勇权郭春明许晓椿
Owner 天津力牧生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap