Method for promoting in-vitro growth of cut and sampled embryo after freezing and unfreezing

A technology of freezing, thawing, cutting and sampling, which is applied in the field of agriculture-animal husbandry and veterinary medicine, can solve problems such as improving the recovery rate, and achieve the effects of improving pregnancy rate, increasing economic and social benefits, and increasing utilization efficiency

Inactive Publication Date: 2011-11-23
CHINA AGRI UNIV
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0004] At present, the research on frozen embryos after cutting and sampling for gender identification mainly focuses on how to reduce the damage caused by the embryo sampling process and freezing process and the length of short-term culture before freezing after cutting. A report on the recovery rate study

Method used

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  • Method for promoting in-vitro growth of cut and sampled embryo after freezing and unfreezing
  • Method for promoting in-vitro growth of cut and sampled embryo after freezing and unfreezing
  • Method for promoting in-vitro growth of cut and sampled embryo after freezing and unfreezing

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Embodiment

[0024] 1. Embryo Production

[0025] 14-17 month-old Holstein bred cattle, normal reproductive system, no reproductive diseases, good body condition, medium body condition. Artificial insemination was performed after synchronous estrus and superovulation, and embryos were punched by non-surgical methods on the 7th day. According to the condition of the cervix of the donor cow, the blasting tube is selected, and a 60mL syringe is directly connected to the blasting tube for embryo punching. Each donor uterine horn uses 220mL PBS egg flushing solution. Filter the recovered solution with an embryo filter, and pour the remaining 30 mL of the recovered solution into a φ90 mm petri dish for microscopic examination.

[0026] 2. Embryo gender identification

[0027] The present invention adopts PCR method to carry out gender identification to dairy cow embryo, and concrete method is as follows:

[0028] ① Calibrate the cutting knife on the micromanipulator.

[0029] ②Pipe the embry...

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Abstract

The invention provides a method for promoting the in-vitro growth of an embryo cut and sampled for sex identification after freezing and unfreezing, which is characterized in that: after freezing and unfreezing the embryo from which a cell sample is cut for sex identification, the embryo is subjected to in-vitro culture in culture solution containing melatonin; the hatching rate of in-vitro culture after the embryo is cut, sampled and frozen is improved; the damage of cutting and freezing to embryo is reduced; the implantation potential after embryo transplantation is increased; and thus, the utilization efficiency of the embryo after sex identification is increased. In addition, the melatonin (MT) used in the method is directly dissolved in the culture solution (for example CR1aa solution) instead of the conventional dimethylsulfoxide (DMSO) and ethanol, so the possible toxic effect of the DMSO and ethanol on the embryo is avoided.

Description

technical field [0001] The invention belongs to the field of agriculture-animal husbandry and veterinary medicine, and in particular relates to a method for promoting in vitro development of cut and sampled embryos after freezing and thawing. Background technique [0002] In 1904, Spemann divided the embryos of frogs and obtained homozygous twin offspring. After that, the researchers carried out experiments on the embryos of mice, rabbits, sheep and other animals at different stages after division, culture and transplantation. By 1988, The first split embryonic mouse in my country completed by Professor Zhang Yong was born in Northwest Agricultural University. At present, there are pigs, horses, cattle, sheep, rabbits, mice, and human split embryos. With the development of embryo biotechnology, embryo segmentation and embryo sex identification technology has become an indispensable part. Embryo gender identification technology has promoted the industrialization of embryo tra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073C12Q1/68
Inventor 刘国世李树静田秀芝高超张洪海
Owner CHINA AGRI UNIV
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