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Method for observing dynamic change of mouse oocytes by using low-melting-point agarose

An oocyte, dynamic change technology, applied in the biological field, can solve the problems of cumbersome process, complicated operation and installation, affecting other people to do experiments, etc., to achieve the effect of solving complexity, low price, and avoiding image loss

Active Publication Date: 2019-12-03
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this device is more complicated to operate and install.
Live cell workstations are generally on public platforms, and installing a perfusion device in a small incubator will affect other people's experiments
If you remove the device after use and install it when you need it, the whole process will be cumbersome

Method used

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  • Method for observing dynamic change of mouse oocytes by using low-melting-point agarose
  • Method for observing dynamic change of mouse oocytes by using low-melting-point agarose
  • Method for observing dynamic change of mouse oocytes by using low-melting-point agarose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Weigh 0.1 g of low-melting point agarose powder and put it into 10 mL of KSOM culture solution, mix well and dissolve in water at 70°C for 2 hours to allow it to fully dissolve.

[0037]Put multiple mouse germinal vesicle oocytes pre-incubated with the pH-sensitive dye BCECF-AM for 20 min into 10 μL of KSOM culture droplets, and increase the contact area between the KSOM culture droplets and the culture dish by slightly shaking the culture dish. Larger, the droplet height becomes shorter. Then, 20 μL of 1% low-melting point agar in a liquid state at 37°C was prepared to cover the KSOM culture drop prepared above by drawing a circular spiral with multiple mouse germinal vesicular oocytes as the center through a relatively thick mouth pipette. Since the KSOM culture solution will dilute 1% low-melting point agarose, its concentration will be reduced to about 0.67%. Cool at room temperature for 10 minutes, and add 37°C KSOM culture solution to the cell culture dish after ...

Embodiment 2

[0042] Weigh 0.1 g of low-melting point agarose powder and put it into 10 mL of KSOM culture solution, mix well and dissolve in water at 70°C for 2 hours to allow it to fully dissolve.

[0043] Put multiple mouse first meiotic metaphase (MI) oocytes that had been pre-incubated for 20 min with the pH-sensitive dye BCECF-AM into 10 μL of KSOM culture droplet, and slightly shake the culture dish to make the KSOM culture droplet and The contact area of ​​the Petri dish increases and the droplet height becomes shorter. Then, 20 μL of 1% low-melting point agar in a liquid state at 37°C was prepared to draw a circular spiral with multiple mouse MI stage oocytes as the center to cover the above-mentioned prepared KSOM culture droplet through a relatively thick mouth pipette. KSOM medium will dilute 1% low-melting point agarose to a concentration of about 0.67%. Cool at room temperature for 10 minutes, and add 37°C KSOM culture solution to the cell culture dish after the agarose gels....

Embodiment 3

[0048] Weigh 0.1 g of low-melting point agarose powder and put it into 10 mL of KSOM culture solution, mix well and dissolve in water at 70°C for 2 hours to allow it to fully dissolve.

[0049] Put multiple mouse second meiosis metaphase (MII) oocytes pre-incubated for 20 min with the pH-sensitive dye BCECF-AM into 10 μL of KSOM culture droplet, and slightly vibrate the culture dish to make the KSOM culture droplet and The contact area of ​​the Petri dish increases and the droplet height becomes shorter. Then, 20 μL of 1% low-melting point agar in a liquid state at 37°C was prepared to draw a circular spiral with multiple mouse MII stage oocytes as the center to cover the prepared KSOM culture droplet above through a relatively thick mouth pipette. KSOM medium will dilute 1% low-melting point agarose to a concentration of about 0.67%. Cool at room temperature for 10 minutes, and add 37°C KSOM culture solution to the cell culture dish after the agarose gels.

[0050] Put the ...

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Abstract

The invention discloses a method for observing dynamic change of mouse oocytes by using a low-melting-point agarose, and belongs to the technical field of biology. The method comprises the following steps: preparing a low-melting-point agarose solution with a mass volume concentration of 1%, covering the low-melting-point agarose solution on a culture liquid drop containing living cells, and cooling at room temperature to obtain an agarose gel containing the living cells for observation in different processing environments. The method has advantages that the characteristics of low-melting-point agarose such as low price, no toxicity, high sieving property, high transparency and the like are fully utilized, and real-time dynamic change observation of a plurality of mouse oocytes under different treatment conditions is realized by the method with simple operation and device.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a method for observing dynamic changes of mouse oocytes by using low-melting point agarose. Background technique [0002] Oocytes are a type of reproductive cells of both sexes. When the paternal sperm fertilizes the maternal egg, the paternal sperm only provides the genetic material DNA, while all other nutrients, energy substances, structural tissues and half of the genetic material DNA of the fertilized egg come from the maternal egg. Therefore, the quality of oocytes has an important impact on the success of fertilization in the later stage, or the quality of fertilized eggs and early embryos after fertilization. In addition, it is widely recognized from demographics and epidemiological pathology that after the age of 35, the quality of oocytes decreases with age. Therefore, it is of great significance to be able to accurately understand the differences in t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12N5/075
CPCG01N33/5005C12N5/0609C12N2500/34
Inventor 程金妹孙斐范晓博
Owner NANTONG UNIVERSITY
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