Culture medium for preserving organoids at room temperature and method for maintaining growth activity of organoids

A technology of organoids and medium, applied in the field of organoid culture, can solve the problems of lack of reports and low growth activity of organoids, and achieve the effect of increasing the number, maintaining growth activity, and improving growth activity

Active Publication Date: 2021-04-20
南昌五元生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are not many reports on the medium formulation of organoids, and there

Method used

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  • Culture medium for preserving organoids at room temperature and method for maintaining growth activity of organoids
  • Culture medium for preserving organoids at room temperature and method for maintaining growth activity of organoids
  • Culture medium for preserving organoids at room temperature and method for maintaining growth activity of organoids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-3

[0052] Embodiment 1-3, a kind of culture medium that can be used for storing organoids at room temperature

[0053] Table 2 shows the composition of the culture medium used in the present invention for storing organoids at room temperature.

[0054] Table 2

[0055]

[0056]

Embodiment 4

[0057] Example 4. A method for maintaining the growth activity of organoids

[0058] A method for maintaining organoid growth activity, comprising the steps of:

[0059] S1. Pretreatment of organoids: Discard the medium of liver organoids in the 24-well plate in a biosafety cabinet, add 900 μl of pre-cooled PBS buffer to each well, and use a 1ml gun to aspirate the pre-cooling in the 24-well plate blow off the matrigel containing organoids, and transfer to a 15ml centrifuge tube, add pre-cooled PBS buffer to make up to 10ml, centrifuge at 333g, 0℃ for 5 minutes; discard the supernatant, Add trypsin to digest into a single cell, add 0.2ml FBS solution to stop the digestion, centrifuge at 333g, 0°C for 5 minutes, discard the supernatant, resuspend the pellet with 500μl medium, dilute the resuspended sample with the expansion medium Organs and organoids are divided into three densities of 100%, 50%, and 25%. Add an equal volume of Matrigel and mix to obtain an organoid suspensio...

Embodiment 5

[0063] Example 5. A method for maintaining the growth activity of organoids

[0064] Same as Example 4, the only difference is that Example 5 does not contain step S3, and other parameters are the same as the preparation method.

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PUM

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Abstract

The invention relates to the technical field of organoid culture, in particular to a culture medium for preserving organoids at room temperature and a method for maintaining the growth activity of the organoids. The culture medium for preserving the organoids at room temperature comprises a DMEM high-glucose culture medium, Glutamax, HEPES, antibiotics, Matrigel, additives, a TGF-beta inhibitor and a Wnt signal pathway activator, and the volume fraction of the Matrigel is 20%-25% in the culture medium for preserving the organoids at room temperature. The culture medium provided by the invention can improve the biological activity of organoid preservation, and the method for maintaining the growth activity of the organoids can ensure that the organoids can maintain the activity for the longest time at room temperature, the growth activity and morphology of the organoids are not influenced, the activity can be quickly recovered after the organoids are transferred into a cell incubator, the growth of the organoids is continued, and the dissociation phenomenon is avoided.

Description

technical field [0001] The invention relates to the technical field of organoid culture, in particular to a medium for storing organoids at room temperature and a method for maintaining growth activity of organoids. Background technique [0002] Organoids are 3D cultured cells with stem potential to form part of the corresponding organ tissue. Organoids can accurately simulate the epithelial structure in vivo and can be subcultured for a long time. The purpose of these models is to illustrate the function of the body from the molecular level down to the cell, tissue, organ or whole organism. At present, organoid models such as human and mouse stomach, small intestine, liver, pancreas, and prostate have been successfully established. These organoid models can be cultured for a long time and have stable phenotypic and genetic characteristics. The culture of organoids can establish reliable disease models and also serve as a new way to screen drugs. The cultivation of organoi...

Claims

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Application Information

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IPC IPC(8): C12N5/00A01N1/02
Inventor 周正牛建军刘平果王栋杨光张以平
Owner 南昌五元生物科技有限公司
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