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Culture liquid for promoting ectogenesis of frozen embryo after thawing

An in vitro development and culture medium technology, applied in the field of agriculture, animal husbandry and veterinary medicine, can solve the problem of increasing the embryo cleavage rate and the number of blastocyst cells, the blastocyst rate is not significantly improved, and the effect of the cleavage rate and the number of blastocyst cells is not significant. and other problems, to achieve the effect of improving late development ability, increasing the number of cells in the embryo, and avoiding toxic effects

Inactive Publication Date: 2012-08-15
定州市中农大朝来农产品开发有限责任公司
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Problems solved by technology

In addition, 68.85% (42 / 61) of the blastocyst development rate of the experimental group added to the in vitro development fluid of bovine embryos was significantly higher than that of the control group (50.79% (32 / 63) (Papis et al., 2007); In the study of cell maturation in vitro and parthenogenetically activated embryo in vitro development, it was also found that adding 10 -9 mol / LMT significantly increased the cleavage rate and blastocyst cell number of embryos, but the blastocyst rate did not increase significantly (N. Rodriguez-Osorio et al., 2007); 10ng / mL MT could significantly increase the polar body The expulsion rate (84.6% vs 75.6%) and the rate of embryo development to blastocyst after parthenogenetic activation, but had no significant effect on cleavage rate and blastocyst cell number (Jung-Teak Kang et al., 2008)

Method used

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  • Culture liquid for promoting ectogenesis of frozen embryo after thawing
  • Culture liquid for promoting ectogenesis of frozen embryo after thawing
  • Culture liquid for promoting ectogenesis of frozen embryo after thawing

Examples

Experimental program
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Effect test

Embodiment

[0023] 1. Superovulation in mice

[0024] 5-6-week-old female Kunming white mice (purchased from the Experimental Animal Center of the Chinese Academy of Military Medical Sciences) had free access to food and water, controlled light (14h / d light), and the indoor temperature was 20-22°C. After a week of adaptive feeding, 10 IU of PMSG and 10 IU of hCG were injected intraperitoneally every other day, and then mated with sexually mature male mice of the same lineage. The vaginal plug was checked the next morning, and female mice with plug were used to obtain 2-cell embryos.

[0025] 2. 2-cell embryo collection

[0026] 48 hours after the injection of hCG, the female mice with tethered were killed by cervical dislocation, the oviducts were cut, and the ampulla of the oviducts was found under a microscope, and the tube wall was cut with the tip of the syringe, and the 2-cell embryos with normal morphology were collected in M 2 Wash in solution (Table 3) and set aside. The operati...

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Abstract

The invention provides a culture liquid for promoting ectogenesis of embryo vitrified by an open pulled straw (OPS) method after thawing. Frozen 2-cell embryo after thawing is cultured in the culture liquid containing 10<-9> mol / L of melatonin (MT), so that the blastula development rate of the embryo is improved and the number of the cells in the embryo is increased, thereby improving the embryo quality, and providing a beneficial reference for further developing the study of OPS cryopreservation of livestock embryos.

Description

technical field [0001] The invention belongs to the field of agriculture-animal husbandry and veterinary medicine, and in particular relates to a culture solution for promoting in vitro development of frozen embryos after thawing. Background technique [0002] Embryo freezing is a practical technique for long-term preservation of animal germplasm resources. The development and application of embryo freezing technology has reduced the limitation of time and space on embryo manipulation, and can effectively avoid the spread of diseases caused by the transportation of live animals. Since the method of vitrification was reported by Rail and Fahy in 1985, this method has received extensive attention and developed rapidly because it does not require expensive freezing equipment, is easy to operate, and has a short freezing process. Vitrification is the use of a high-concentration cryoprotectant to form a glassy state during rapid cooling, effectively avoiding the formation of har...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/073
Inventor 刘国世朱士恩田秀芝史文清张洪海
Owner 定州市中农大朝来农产品开发有限责任公司
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