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Culture medium for epitaxy culture of mouse embryo body

A mouse embryo and culture medium technology, applied in the field of in vitro culture of mammalian embryos, can solve the problems of inability to meet the needs of in vitro embryos, influence, and low success rate

Pending Publication Date: 2022-04-15
SHANXI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The existing in vitro embryo culture technology can only cultivate mouse embryos up to E6.5, which cannot meet the needs of in vitro embryo experiments, and the success rate is low, only 36.7%, which may affect subsequent research in actual experiments

Method used

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  • Culture medium for epitaxy culture of mouse embryo body
  • Culture medium for epitaxy culture of mouse embryo body

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Configuration of culture medium.

[0028] Configure the first culture medium according to the following components:

[0029] Add 1 mL of fetal bovine serum, 1 mL of human umbilical cord serum, 100 μL of sodium pyruvate at a concentration of 100 mM, 100 μL of glutamine at a concentration of 200 mM, 100 μL of penicillin-streptomycin at a concentration of 10,000 units, and 100 μL of 100× N2 additive and 200 μL B27 additive with a concentration of 50×, mixed evenly and stored at 4°C until use.

[0030] Configure the second culture medium after culturing for 48 h according to the following components:

[0031] Add 2 mL of fetal bovine serum, 1 mL of human umbilical cord serum, 100 μL of sodium pyruvate at a concentration of 100 mM, 100 μL of glutamine at a concentration of 200 mM, 100 μL of penicillin-streptomycin at a concentration of 10,000 units, and 100 μL of 100× N2 additive and 200 μL B27 additive with a concentration of 50×, mixed evenly and stored at 4°C ...

Embodiment 2

[0032] Example 2 In vitro culture of mouse embryos.

[0033] Culture mouse embryos as follows:

[0034] 1) Extract E3.5 mouse blastocysts from the uterus of 6-7-week-old pregnant female ICR mice, a total of 25, wash the blastocysts in fresh and preheated M2 medium for 3-5 times, and wash with glass Transfer the tube to 30 μL of acidic Tyrode’s solution micro-droplet. After the zona pellucida is digested, transfer it to the micro-droplet of 30 μL fresh M2 medium immediately. Mix the blastocysts after digesting the zona pellucida with 100 μL of 30% volume concentration Matrigel was mixed, and inoculated onto a 24-well culture plate pre-coated with extracellular matrix gel (100 μL / well), so that the glue wrapped the embryos, and about 10 embryos were inoculated in each well to establish a 3D in vitro culture platform;

[0035] 2) Add 500 μL of the first medium prepared in Example 1 to each well of the culture plate, and after culturing for 48 hours, replace and use the second me...

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Abstract

The invention discloses a culture medium for epitaxy culture of a mouse embryo body, which contains a basic culture medium, fetal calf serum, human umbilical serum, sodium pyruvate, glutamine, penicillin-streptomycin, an N2 additive and a B27 additive, and can promote the continuous culture stage of an in-vitro mouse embryo to E8.0 on the basis of improving the culture success rate. A foundation is provided for development research of pre-implantation embryos and implantation stage embryos, the selection range of mouse embryo related experiment contents and experiment stages can be expanded by improving the culture success rate and prolonging the in-vitro culture time, and a basis can also be provided for in-vitro culture of other mammal embryos.

Description

technical field [0001] The invention relates to the field of in vitro culture of mammalian embryos, in particular to a culture medium for in vitro time-delayed culture of mouse embryos. Background technique [0002] Mouse embryo is one of the most commonly used mammalian embryo models, and it has become a good model for embryo research due to its advantages of mammalian embryonic characteristics and short development time. [0003] The establishment of an embryo continuous culture system in vitro is an important platform to elucidate the developmental mechanism of mammalian embryos. At present, there are some research results on the preimplantation embryonic development of mice, but the research on the implantation period and postimplantation period is very limited. The main reason is that the survival rate of in vitro embryos is not high, there is no promotion of various factors in the body, and the culture time of embryos in vitro is also very short. Moreover, the method o...

Claims

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Application Information

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IPC IPC(8): C12N5/073
CPCC12N5/0604C12N2500/84C12N2500/32C12N2500/30C12N2513/00
Inventor 解军梁宇翔金姗姗王莹丰子涵闫瞻遥吕慧敏张潇刘志贞侯淑玲赵虹奥瑞芳李航王磊杨丽红
Owner SHANXI MEDICAL UNIV
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