A kind of mouse embryo transplant glue and preparation method thereof

A mouse embryo, mouse technology, applied in the field of reproduction, can solve the problems of embryo deformation, low implantation rate of embryo transfer, and unstable development.

Active Publication Date: 2021-08-20
FUYANG NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During embryo transfer in mice, the embryo will be deformed rapidly and the implantation rate of the embryo transfer is low, and the development of the embryo after transfer is unstable, resulting in a high rate of miscarriage

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The present embodiment is a mouse embryo transplant glue, the specific preparation process is as follows:

[0023] The amniotic membrane after childbirth is used as the raw material, and the decellularized amniotic membrane is obtained after the decellularization treatment with trypsin combined with Triton X-100; under the condition of liquid nitrogen freezing, the obtained decellularized amniotic membrane is subjected to high-speed grinding to obtain granular fragments, And sterilize the obtained granular fragments with cobalt 60 irradiation to obtain granular amniotic membrane; inoculate 1.0×10 per 1 mg of granular amniotic membrane 5 The ratio of umbilical cord mesenchymal stem cells, umbilical cord mesenchymal stem cells were inoculated on the granular amniotic membrane, and co-cultured for 5 hours to obtain a stem cell-granular amniotic membrane mixture. ; Take fresh blood samples isolated from mice, obtain platelet-rich plasma by centrifugation, store at 4°C for l...

Embodiment 2

[0025] In this example, a mouse embryo transplant glue, the specific preparation process is the same as in Example 1, the only difference is that the amount of each substance added in each mL of HEPES buffer is as follows: 0.05 μg of pregnant horse serum, 0.05 μg of chorionic gonadotropin 375 units of hormone, 50 μg of streptomycin, 20 μL of amniotic substance, and 10 μL of mouse plasma.

Embodiment 3

[0027] In this example, a mouse embryo transplant glue, the specific preparation process is the same as in Example 1, the only difference is that the amount of each substance added in each mL of HEPES buffer is as follows: 0.035 μg of pregnant horse serum, 0.035 μg of chorionic gonadotropin 100 units of hormone, 42 μg of streptomycin, 20 μL of amniotic substance, and 8 μL of mouse plasma.

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Abstract

The invention discloses a mouse embryo transplant glue and a preparation method thereof. The mouse embryo transplant glue is prepared by adding pregnant horse serum, chorionic gonadotropin, amniotic membrane substances, mouse plasma, and streptomycin to a pH of 7.0 ~7.2, prepared in HEPES buffer solution with a concentration of 20mmol / L; the addition amount of each substance in each mL of HEPES buffer solution is as follows: 0.01~0.05μg of pregnant horse serum, 100~375 units of chorionic gonadotropin, streptomyces 50-70 μg of prime, 20 μL of amniotic substance, and 8-10 μL of mouse plasma. The mouse embryo transplantation glue provided by the invention can not only adhere the embryo to the uterus, improve the implantation rate of the embryo, but also provide nutrition for the growth of the embryo after implantation, and greatly improve the survival rate of the mouse embryo.

Description

technical field [0001] The invention belongs to the technical field of reproduction, and in particular relates to a glue for mouse embryo transplantation and a preparation method thereof. Background technique [0002] Mammalian embryo transfer technology is an important technology in experimental embryology, genetics, and developmental biology research. Embryo transfer is to take out the early embryos of a female animal from the oviduct or uterus and transplant them into the oviduct of another female animal. Or in the uterus, allowing it to continue to develop into a fetus. Mice are the most important model mammals in embryo transfer technology, and the early embryos of mice are commonly used experimental materials for studying the early development mechanism of mammals and humans. Mouse embryo transfer, that is, embryos obtained by in vitro fertilization or other methods are transferred to pseudopregnant recipients, so that the embryos develop in vivo and give birth to off...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/073A61D19/04C12N5/0775
CPCA61D19/04C12N5/0604C12N5/0665C12N2500/84C12N2501/31C12N2533/92
Inventor 刘勇王文英李文雍辛晶孔凤吴巧琴吴晓庆
Owner FUYANG NORMAL UNIVERSITY
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