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Segmentation liquid for bovine embryo segmentation and bovine embryo segmentation method

A technique for bovine embryos and embryos, which is applied to the application of the test solution used for bovine embryo segmentation in bovine embryo segmentation.

Inactive Publication Date: 2021-07-23
天津力牧生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Zheng Haiying’s literature (Zheng Haiying, et al., Effects of different segmentation fluids on the segmentation effect of buffalo embryos, Animal Husbandry and Veterinary Medicine, 2009, 41 (7): 32) has recorded methods for segmenting bovine embryos, but these methods still have certain limitations

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] Embodiment 1: the division method of bovine in vitro fertilization embryo (collecting ovum in living body)

[0099] Cattle used in the experiment: Chinese Yellow Cattle (Nanyang Cattle, a working breed).

[0100] Step (1), collection and in vitro maturation of oocytes

[0101] Live collection: live bovine eggs are collected, and the resulting follicular fluid is picked out under a stereomicroscope to pick out cumulus-oocyte complexes (COCs) that contain at least 3 layers of cumulus cells, and put them into the mature culture medium containing HEPES Temperature 38.8 ℃, transported back to the laboratory within 3 hours;

[0102] The COCs collected above were washed once in the oocyte maturation medium, and then transferred to a new maturation medium for 22–24 h. The culture conditions were 38.8°C, 5.5–6.5% CO2, and saturated humidity, so that the oocyte in vitro maturation;

[0103] Step (2), in vitro fertilization

[0104] Wash the mature COCs once in the fertilizati...

Embodiment 2

[0114] Embodiment 2: the segmentation method of bovine in vitro fertilization embryo (collecting ovum in living body)

[0115] Cattle used in the test: Holstein cattle (dairy cow breed).

[0116] Step (1), collection of oocytes and in vitro maturation

[0117] Live collection: live bovine eggs are collected, and the resulting follicular fluid is picked out under a stereomicroscope to pick out cumulus-oocyte complexes (COCs) that contain at least 3 layers of cumulus cells, and put them into the mature culture medium containing HEPES Temperature 38.8 ℃, transported back to the laboratory within 3 hours;

[0118] The COCs collected above were washed once in the oocyte maturation medium, and then transferred to a new maturation medium for 22–24 h. The culture conditions were 38.8°C, 5.5–6.5% CO2, and saturated humidity, so that the oocyte in vitro maturation;

[0119] Step (2), in vitro fertilization

[0120] Wash the mature COCs once in the fertilization medium, then transfer...

Embodiment 3

[0130] Embodiment 3: the segmentation method of bovine in vitro fertilization embryo (collecting ovum in living body)

[0131] Cattle used in the experiment: Simmental cattle (beef breed).

[0132] Step (1), collection and in vitro maturation of oocytes

[0133] Live collection: live bovine eggs are collected, and the resulting follicular fluid is picked out under a stereomicroscope to pick out cumulus-oocyte complexes (COCs) that contain at least 3 layers of cumulus cells, and put them into the mature culture medium containing HEPES Temperature 38.8 ℃, transported back to the laboratory within 3 hours;

[0134] The COCs collected above were washed once in the oocyte maturation medium, and then transferred to a new maturation medium for 22–24 h. The culture conditions were 38.8°C, 5.5–6.5% CO2, and saturated humidity, so that the oocyte in vitro maturation;

[0135] Step (2), in vitro fertilization

[0136] Wash the mature COCs once in the fertilization medium, then transf...

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Abstract

The invention relates to a segmentation liquid for bovine embryo segmentation and a bovine embryo segmentation method. The segmentation liquid is prepared from 9.39 g of dipotassium phosphate, 3.5 g of monopotassium phosphate, 68 g of cane sugar, 12 g of povidone K30 and the balance of water for injection, wherein the total volume is 1000 ml. The bovine embryo segmentation method comprises the following steps: collecting oocytes and carrying out in-vitro maturation on the oocytes; incubating sperm suspension and the mature oocytes to complete in-vitro fertilization; culturing fertilized embryos to obtain mulberry embryos, and picking out the mulberry embryos; transferring the mulberry embryo into a segmentation liquid drop, and placing the segmentation liquid drop under an inverted microscope to segment the embryo into two parts; and transferring the segmented half embryos into an embryo culture solution, and culturing until the mulberry embryos develop into expanded blastocysts which can be used for embryo transplantation. By using the embryo segmentation liquid in the invention, the embryo segmentation effect can be effectively improved.

Description

technical field [0001] The invention belongs to the technical field of animal reproduction, and relates to a technology for agricultural-animal husbandry and veterinary reproduction, in particular to a method for dividing bovine embryos in vitro. In addition, the present invention also relates to the use of test solutions for the division of bovine embryos in the division of bovine embryos. application. The bovine embryo segmentation method of the invention has excellent technical effects. In particular, the present invention relates to a technique for segmenting embryos obtained after in vitro fertilization of bovine oocytes obtained from in vitro fertilization, and the embryo segmentation fluid used when performing this embryo segmentation technique, using the Embryo segmentation fluid can effectively improve the effect of embryo segmentation. Background technique [0002] With the development of modern agricultural science and technology, in order to make full use of th...

Claims

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Application Information

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IPC IPC(8): C12N5/073
CPCC12N5/0604C12N2500/12C12N2500/14C12N2500/16C12N2500/24C12N2500/30C12N2500/32C12N2500/33C12N2500/34C12N2500/35C12N2500/36C12N2500/38C12N2500/40C12N2500/60C12N2501/105C12N2501/11C12N2501/31C12N2501/392C12N2501/91C12N2501/998C12N2501/999C12N2509/00C12N2509/10C12N2517/10
Inventor 王小武韩勇权王娜陈建美郭春明许晓椿
Owner 天津力牧生物科技有限公司
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