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Culture solution capable of improving in vitro maturation rate of pig oocytes and application thereof

An oocyte, in vitro maturation technology, applied in the field of pig genetics and breeding, can solve the problems of low efficiency and lack of high-quality mature oocytes

Inactive Publication Date: 2017-08-29
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the maturity rate of porcine oocytes cultured in vitro in domestic laboratories is generally 60-70%, and the efficiency is still relatively low, so that the implementation of somatic cell nuclear transfer technology is limited by the lack of high-quality mature oocytes

Method used

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  • Culture solution capable of improving in vitro maturation rate of pig oocytes and application thereof
  • Culture solution capable of improving in vitro maturation rate of pig oocytes and application thereof
  • Culture solution capable of improving in vitro maturation rate of pig oocytes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] 1 material

[0016] TCM-199, NaHCO3, D-glucose, Sodium Pyruvate, Streptomycin sulfate, PVA, Cysteine, Penicillin G, EGF, FBS, HCG, PMSG, Milli-Q water, egg wash.

[0017] 2 Methods and results

[0018] 2.1 Collection of follicular fluid

[0019] Sow ovaries were collected from slaughterhouses in Nanning City, placed in normal saline at 34°C for 2 hours, and transported back to the laboratory, and washed three times with 0.9% double-antibody normal saline. The follicular fluid in pig follicles with a diameter of 3-6 mm was extracted with a syringe, centrifuged at 1600 g / min for 20 min, the supernatant was taken, filtered with a 0.22 μm filter, and then packed in aliquots, and stored at -20°C until use.

[0020] 2.2 Preparation of maturation solution

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Abstract

The invention belongs to the technical field of pig genetic breeding, and specifically relates to a culture solution capable of improving in vitro maturation rate of pig oocytes and application thereof. The culture solution capable of improving the in vitro maturation rate of pig oocytes is prepared from the following raw materials with the following concentrations: 9.5 g / L of TCM-199, 2.2 mg / mL of NaHCO3, 3.05 mmol / L of D-glucose, 0.91 mmol / L of Sodium Pyruvate, 0.5 g / L of Streptomycin sulfate, 0.05% of PVA, 0.57 mmol / L of Cysteine and 0.075 g / L of Penicillin G. The 0.05% of PVA and 5% of a follicular fluid are added on the basis of pig oocyte in vitro maturation culture solution widely used in our country, so that the in vitro maturation rate of the pig oocytes can be improved to more than 90%, and matured oocytes with higher quality and more quantity can be provided for porcine somatic cell nuclear transfer technology.

Description

technical field [0001] The invention belongs to the technical field of pig genetics and breeding, and in particular relates to a culture medium for improving the in vitro maturation rate of pig oocytes and its application. Background technique [0002] Porcine somatic cell nuclear transfer cloning technology is currently the easiest and most reliable method to construct transgenic pigs. Among the many factors that affect the efficiency of porcine nuclear transfer, the quality of oocytes takes the first place. At present, the maturation rate of pig oocytes cultured in vitro in domestic laboratories is generally 60-70%, and the efficiency is still relatively low. The implementation of somatic cell nuclear transfer technology is limited by the lack of high-quality mature oocytes. [0003] Follicular fluid and polyvinyl alcohol (PVA) are the two most commonly used ingredients for oocyte maturation culture medium. Due to the complex composition of follicular fluid and a variety ...

Claims

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Application Information

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IPC IPC(8): C12N5/075
CPCC12N5/0609C12N2500/12C12N2500/32C12N2500/34C12N2500/35
Inventor 张笠蒋钦杨郭亚芬兰干球
Owner GUANGXI UNIV
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