Method for improving cultivation efficiency of bovine in vitro fertilization embryos

An embryo culture medium, bovine body technology, applied in embryo cells, culture process, tissue culture and other directions, can solve the problems of low efficiency, high cost of in vitro fertilization embryo production, and limited wide application.

Active Publication Date: 2017-09-08
无锡博裕力牧生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Although in vitro fertilization technology can be successfully applied to many mammals, the high cost and low efficiency of in vitro fertilized embryo production due to the low blastocyst rate of in vitro fertilization limit the wide application of this technology in the practice of rapid breeding of cattle

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0145] Embodiment 1: the culture method of bovine in vitro fertilization embryo

[0146] 1. Reagents

[0147] Mature medium: Containing 0.01IU / ml FSH, 10IU / ml LH, 1μg / ml estradiol, 100ng / m l TCM-199 culture solution of IGF, 50ng / ml EGF, 100U / ml penicillin, 100μg / ml streptomycin, 10% fetal bovine serum.

[0148] Wash semen: Containing 112.0mM NaCl, 4.02mM KCl, 2.25mM CaCl2 2H2O, 0.52mM MgCl2 6H2O, 0.83mM KH2PO3, 37.0mM NaHCO3, 1.25mM sodium pyruvate, 10μg / ml heparin, 4mg / ml of bovine serum albumin (Bovine serum albumin, BSA), 10 mM caffeine, 100 U / ml of penicillin, and 100 μg / ml of streptomycin in water.

[0149] Semen received: Containing 112.0mM NaCl, 4.02mM KCl, 2.25mM CaCl2 2H2O, 0.52mM MgCl2 6H2O, 0.83mM KH2PO3, 37.0mM NaHCO3, 1.25mM sodium pyruvate, 10μg / ml heparin, 4mg / ml of BSA, 100 U / ml of penicillin, and 100 μg / ml of streptomycin in water.

[0150] Pre-embryo culture medium: Contains 109.5mM NaCl, 3.1mM KCl, 26.2mM NaHCO3, 0.8mM MgCl2 6H2O, 1.19mM KH2...

Embodiment 2

[0201] Embodiment 2: the culture method of bovine in vitro fertilization embryo

[0202] With reference to Example 1, the only difference is that the composition of the embryonic late stage culture solution is changed to:

[0203] 109mM NaCl,

[0204] 3.1mM KCl,

[0205] 26.0mM NaHCO3,

[0206] 0.5mM MgCl2 6H2O,

[0207] 1.3mM KH2PO3,

[0208] 0.4mM sodium pyruvate,

[0209] 1.5mM glucose,

[0210] 5mM Calcium Galactonate,

[0211] 10v / v% fetal bovine serum,

[0212] 1mM L-Glutamine,

[0213] 2v / v% essential amino acids,

[0214] 1v / v% non-essential amino acids,

[0215] 3mM glutathione, and

[0216] Water as dosing solvent.

[0217] The test was carried out as in Example 1, and all the indicators of the results were basically the same as those of the 3mM glutathione group in Example 1. For example, in this example, the morula rate was 0.633, the blastocyst rate was 0.508, and the apoptosis rate was 0.050.

Embodiment 3

[0218] Embodiment 3: the culture method of bovine in vitro fertilization embryo

[0219] With reference to Example 1, the only difference is that the composition of the embryonic late stage culture solution is changed to:

[0220] 110mM NaCl,

[0221] 2.9mM KCl,

[0222] 26.5mM NaHCO3,

[0223] 1.0mM MgCl2 6H2O,

[0224] 1.0mM KH2PO3,

[0225] 0.4mM sodium pyruvate,

[0226] 1.5mM glucose,

[0227] 5mM Calcium Galactonate,

[0228] 10v / v% fetal bovine serum,

[0229] 1mM L-Glutamine,

[0230] 2v / v% essential amino acids,

[0231] 1v / v% non-essential amino acids,

[0232] 3mM glutathione, and

[0233] Water as dosing solvent.

[0234] The test was carried out as in Example 1, and all the indicators of the results were basically the same as those of the 3mM glutathione group in Example 1. For example, in this example, the morula rate was 0.636, the blastocyst rate was 0.521, and the apoptosis rate was 0.050.

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PUM

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Abstract

The invention relates to a method for improving the cultivation efficiency of bovine in vitro fertilization embryos. In particular, the invention relates to bovine in vitro fertilization embryo cultivation liquid. The cultivation liquid contains NaCl, KCl, NaHCO3, MgCl2, KH2PO3, sodium pyruvate, glucose, calcium galactonate, fetal calf serum, L-glutamine, necessary amino acid, unnecessary amino acid, glutathione and water. The invention also relates to a method for improving the cultivation efficiency of the bovine in vitro fertilization embryos. The method comprises the following steps: putting the bovine in vitro fertilization embryos into the bovine in the vitro fertilization embryo cultivation liquid, and performing embryo in vitro cultivation under the conditions of 38.5 DEG C, 0.5 percent CO2 and 100 percent humidity. The method provided by the invention and the used cultivation liquid present excellent technical effects as stated in the specification.

Description

technical field [0001] The invention belongs to the technical field of animal reproduction, relates to a technology for agriculture-animal husbandry and veterinary reproduction, in particular to a method for improving the culture efficiency of bovine in vitro fertilized embryos, more specifically, the present invention relates to the use of bovine in vitro fertilized embryo culture fluid in Application in bovine in vitro fertilization embryo culture. Further, the present invention also relates to a method for culturing bovine IVF embryos using the bovine IVF embryo culture solution. In particular, the method for culturing bovine IVF embryos of the present invention has excellent cost advantages. 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 IV...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073
CPCC12N5/0603C12N2500/12C12N2500/16C12N2500/30C12N2500/32C12N2500/34C12N2501/999
Inventor 许晓椿王小武郭晶陈敏
Owner 无锡博裕力牧生物科技有限公司
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