Low-temperature preservation solution and preservation method thereof

A low-temperature preservation and preservation method technology, which is applied to the preservation, application, and animal husbandry of human or animal bodies, and can solve problems such as high cost, high hardware requirements, and complicated transportation procedures

Active Publication Date: 2021-04-23
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

The invention solves technical problems such as high hardware requirements, complicated transportation proce

Method used

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  • Low-temperature preservation solution and preservation method thereof
  • Low-temperature preservation solution and preservation method thereof
  • Low-temperature preservation solution and preservation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-7

[0049] Embodiment 1-7, cryopreservation solution of the present invention

[0050] The formulations of the cryopreservation solutions of Examples 1-7 of the present invention are shown in Table 3 and Table 4 below.

[0051] table 3

[0052]

[0053]

[0054] Table 4

[0055]

Embodiment 8

[0056] Example 8. A method for preserving organoids

[0057] A method for organoid preservation, comprising the steps of:

[0058] (1) Discard the culture medium of the organoids in the 24-well plate in a biological safety cabinet, and add 900 μl of pre-cooled PBS buffer to each well;

[0059] (2) Use a 1ml pipette gun to suck up the pre-cooled PBS buffer in the 24-well plate to blow off the matrigel containing organoids, transfer each 2 wells to a 15ml centrifuge tube, add the pre-cooled PBS buffer to the total The volume is 10ml;

[0060] (3) Place the centrifuge tube in step (2) at 333g and centrifuge for 5 minutes at a temperature of 0°C;

[0061] (4) Absorb and discard the supernatant, respectively use a pipette to absorb the cryopreservation solution prepared in Examples 1-7 and blow and beat the precipitate repeatedly until the organoids are blown into fragments;

[0062] (5) Add the pre-cooled cryopreservation solution of Example 1-7 to 13ml, cover and mix by invert...

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Abstract

The invention relates to the technical field of cell preservation and transportation, and particularly discloses a low-temperature preservation solution and a preservation method thereof. The low-temperature preservation solution is prepared from a DMEM high-glucose culture medium, Glutamax, HEPES and Penicilline-Streptomycin. The organoids transported by the cryopreservation solution at low temperature do not need to be resuscitated, and can be quickly cultured, is relatively low in cost and relatively high in culture success rate, and the activity of cells can be maintained for a long time. According to the invention, the technical problems of high hardware requirements, complex transportation procedures, high cost, low efficiency, low success rate of cryopreservation recovery, worry about cell growth conditions and the like are solved.

Description

technical field [0001] The invention relates to the technical field of cell preservation and transportation, in particular to a cryopreservation solution and a preservation method thereof. Background technique [0002] Organoids are three-dimensional cultures of cells in vitro that contain some of the key properties of the organ of origin. Organoids are three-dimensional microscopic structures obtained by in vitro 3D culture of cells or tissues with the potential of adult stem cells. [0003] At present, there is no clear method for the transport of organoids, and most of them are transported in the form of ordinary cells. There are two main methods for cell transportation: (1) transportation by liquid nitrogen or dry ice cryopreservation. In this method, the cells are transported in a plastic foam box filled with dry ice or liquid nitrogen, which can generally be stored for 7-8 days. (2) Transport of live cells. This method is suitable for cells in the early or middle l...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 周正牛建军刘平果罗伙军黄静杨光张以平
Owner 南昌五元生物科技有限公司
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