Method for producing in vitro embryos using lamb superovulated oocytes

A technology of oocytes and superovulation, which is applied in the field of producing embryos in vitro by using oocytes in the ovaries of lambs after superovulation, which can solve the problems that restrict the promotion and breeding of fine varieties, achieve high embryo production efficiency and shorten the generation interval , Improving the speed of genetic improvement and the effect of production efficiency

Inactive Publication Date: 2011-12-28
XINJIANG ACADEMY OF AGRI & RECLAMATION SCI +1
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

However, in actual production, some excellent varieties are often affected by various factors during the promotion and breeding process. One is the lack of embryo donors or sources of excellent varieties, and the other is the bottleneck of the embryo production platform or carrier, which will restrict new ones. Promotion and breeding of fine varieties

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Collection of oocytes: Select 4-10 weeks old presexual ewe lambs. Four consecutive days before collection, FSH was injected 40 mg each time, and 500 IU PMSG was injected at the same time for the last time. The ovaries were exposed and fixed by surgery. Use a 10ml syringe containing egg collection solution (M199 stock solution + 25mmol / L HEPES + 0.2% BSA (fetal bovine serum)) to aspirate the follicles on the ovary. After suction, place the liquid containing COCs in the egg collection solution, and count the eggs number of mother cells.

[0027] Preparation of maturation solution: Add 10 μg / mL FSH (follicle-stimulating hormone), 10 μg / mL LH (luteinizing hormone), 1 μg / mL E2 (estrogen), 0.1 mmol / L β-mercaptoethanol to the stock solution of M199 medium, and press Add 20% ESS (estrous ewe serum) to the volume and equilibrate in the incubator for 2 hours.

[0028] Oocyte culture: the culture solution was made into 50 μL / drop of maturation liquid droplets, the collected COCs...

Embodiment 2

[0034] Example 2: Collection of oocytes: 4-10 weeks old presexual ewe lambs were selected. Two consecutive days before the collection, FSH was injected twice, 50 mg each time, and 400 IU PMSG was injected at the same time for the last time. The ovaries were exposed and fixed by surgery. Use a 10ml syringe containing egg collection solution (M199 stock solution + 20mmol / L HEPES + 0.3% BSA (fetal bovine serum)) to aspirate the follicles on the ovary. After suction, place the liquid containing COCs in the egg collection solution, and count the eggs number of mother cells.

[0035]Preparation of maturation solution: Add 8 μg / mL FSH (follicle-stimulating hormone), 12 μg / mL LH (luteinizing hormone), 1.5 μg / mL E2 (estrogen), 0.15 mmol / L β-mercaptoethanol to M199 medium stock solution, and Add 20% ESS (estrous ewe serum) by volume, make a 40 μL / drop mature solution micro-droplet before use, and equilibrate in the incubator for 1 hour.

[0036] Oocyte culture: After the collected COC...

Embodiment 3

[0042] Example 3: Collection of oocytes: 4-10 weeks old presexual ewe lambs were selected. Four consecutive days before collection, FSH was injected 50 mg each time, and 500 IU PMSG was injected at the same time for the last time. The ovaries were exposed and fixed by surgery. Use a 10ml syringe containing egg collection solution (M199 stock solution + 30mmol / L HEPES + 0.2% BSA (fetal bovine serum)) to aspirate the follicles on the ovary, and place the COCs-containing liquid in the egg collection solution after suction.

[0043] Preparation of maturation solution: Add 12 μg / mL FSH (follicle-stimulating hormone), 8 μg / mL LH (luteinizing hormone), 1.5 μg / mL E2 (estrogen), 0.15 mmol / L β-mercaptoethanol to the M199 medium stock solution, and Add 20% ESS (estrous ewe serum) by volume, make 60 μL / drop mature solution micro-droplets before use, and equilibrate in the incubator for 3 hours.

[0044] Oocyte culture: Wash the collected COCs twice in the maturation solution first, trans...

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Abstract

The invention discloses an in vitro embryo production method by lamb superovulation oocytes. The production method provided by the invention is accomplished by the following technical scheme: carrying out in vitro culture on oocytes acquired from lamb body in a maturation medium and maturing into ova, carrying out in vitro fertilization and culture so as to make the ova grow into embryos, and transplanting survived embryos into a surrogate ewe body for breeding. In comparison with the prior art, the method provided by the invention is utilized to adopt the oocytes acquired after the lamb superovulation treatment for the in vitro embryo production and has advantages of large quantity of the acquired oocytes, high production efficiency of embryo and short time for breeding lamb offspring. Therefore, the method provided by the invention has an edge in the application of large scale in vitro embryo production.

Description

technical field [0001] The invention relates to a method for producing embryos in vitro by using oocytes in lamb ovaries, in particular to a method for producing embryos in vitro by using oocytes in ovaries in lambs after superovulation. Background technique [0002] In current animal husbandry production, the breeding of new varieties determines the level of economic benefits. However, in actual production, some excellent varieties are often affected by various factors during the promotion and breeding process. One is the lack of embryo donors or sources of excellent varieties, and the other is the bottleneck of the embryo production platform or carrier, which will restrict new ones. Promotion and breeding of fine varieties. [0003] Therefore, a method for producing embryos in vitro by using oocytes in lamb ovaries, especially a method for producing embryos in vitro by using oocytes in lamb ovaries after superovulation has emerged. Contents of the invention [0004] Th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073
Inventor 万鹏程石国庆倪建宏石文艳郭洪代蓉刘长彬张宾
Owner XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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