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Method for lamb oocyte in-vitro fertilization culture

An in vitro fertilization and oocyte technology, applied in the field of biology, can solve problems such as decreased fertilization effect and polyspermia

Inactive Publication Date: 2016-01-20
YANGZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In vitro fertilization generally adopts the droplet culture method, in which the density of sperm is generally controlled at 2-10×10 6 A / mL, if the sperm density is too low, the fertilization effect will decrease, while if the sperm density is too high, there will be polyspermy into the egg

Method used

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  • Method for lamb oocyte in-vitro fertilization culture
  • Method for lamb oocyte in-vitro fertilization culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0014] 1. Test material

[0015] Fresh ovaries were collected from healthy 1-2-year-old adult ewes and 1-2-month-old lambs of local white goat breeds in Jiangsu.

[0016] BO liquid, washing liquid: weigh 55.0 mg of NaCl6, NaHCO 3 310.4mg, glucose 251.8mg, KCl30.0mg, CaCl 2 16.7 mg, NaH 2 PO 4 11.5 mg, MgCl 2 10.6mg, sodium pyruvate 5.5mg, 1% phenol red 4.00ml, 100IU / ml penicillin, 100IU / ml streptomycin, add ultrapure water to 100ml constant volume, 0.22μm filter filter sterilization for use.

[0017] BO capacitation solution, BO semen solution (prepared and ready to use): BO solution, heparin sodium 20 μg / mL, BSA 10 mg / mL, mixed and filtered through a 0.22 μm filter to sterilize, 38.5 ° C CO 2 Equilibrate in the incubator for 30 minutes.

[0018] Fertilized egg culture medium: TCM199, FBS20%, NaHCO 3 2.2mg / mL, sodium pyruvate 25.0mg / mL, HEPES6.0mg / mL, sodium penicillin and streptomycin sulfate 100IU / ml each. ,

[0019] 2. Oocyte Collection

[0020] After the ovaries ...

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Abstract

The invention belongs to the field of biology, and particularly relates to a method for lamb oocyte in-vitro fertilization culture. The method comprises the steps of oocyte acquisition and maturation culture, sperm capacitation, mature oocyte in-vitro fertilization and zygote culture. During sperm capacitation and mature oocyte in-vitro fertilization, incubating time, the rotating speed of a centrifugal machine, centrifugation time, the number of times of centrifugal washing and mature oocyte culture time conditions are optimized, so that the zygote cleavage rate of lambs is basically the same as that of adult ewes.

Description

technical field [0001] The invention belongs to the field of biology, and in particular relates to a method suitable for in vitro fertilization and culture of lamb oocytes. Background technique [0002] Juvenile invitroembryotransfer technology (juvenile invitroembryotransfer, JIVET) integrates a series of reproductive technologies such as superovulation technology of young dams, oocyte in vitro maturation technology, in vitro fertilization technology, embryo in vitro culture technology and embryo transfer technology. biotechnology system. JIVET technology uses a large number of oocytes on the ovaries of young female animals as egg sources to culture and produce embryos in vitro, which is conducive to shortening the generation interval of livestock reproduction, fully exploring the reproductive potential of female animals with excellent genetic traits, and improving the speed of genetic improvement and production of herds. Efficiency, providing abundant genetic resources fo...

Claims

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Application Information

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IPC IPC(8): C12N5/073C12N5/075C12N5/076
Inventor 李拥军蔡缪荧程国虎张昊郭海燕
Owner YANGZHOU UNIV
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