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Hepatocyte ultrasonic planting and freezing storage device and freezing storage method thereof

A technology of ultrasound and hepatocytes, which is applied in the preservation, application, and animal husbandry of human or animal body, and can solve the problems of less research and application.

Pending Publication Date: 2021-04-20
UNIV OF SHANGHAI FOR SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few researches and applications on ultrasound-induced ice crystal formation

Method used

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  • Hepatocyte ultrasonic planting and freezing storage device and freezing storage method thereof
  • Hepatocyte ultrasonic planting and freezing storage device and freezing storage method thereof
  • Hepatocyte ultrasonic planting and freezing storage device and freezing storage method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1: experimental method

[0063] 1.1 Materials and reagents

[0064] L-02 primary liver cells (purchased from Shanghai Fuheng Biotechnology Co., Ltd.), fetal bovine serum (Gemini Company), medium RPMI-1640 (Gibco Company), trypsin (TBD Company), dimethyl sulfoxide Me2SO ( Aladdin Company), PBS buffer solution (TBD Company), ethylene glycol (McLean), T-25 culture bottle (NEST Nice), 50ml centrifuge tube (NEST Nice), 15ml centrifuge tube (NEST Nice), 1.5ml sterile centrifuge tube (Axygen), 2ml cryopreservation tube (NEST), AO / PI double staining kit (BestBio).

[0065] 1.2 Instruments and equipment

[0066] Optical microscope (Nikon, Japan), fluorescence microscope (Nikon, Japan), low-speed desktop centrifuge (Shanghai Anting), CO2 cell incubator (Shanghai Boxun), programmed cooling box (Thermo Fisher Biochemical Co., Ltd. ), ultra-low temperature refrigerator (Qingdao Haier Special Electric Co., Ltd.), low-temperature constant temperature bath (Ningbo Tianhen...

Embodiment 2

[0100] Example 2: Results and Discussion

[0101] 2.1 Establishment of the upper threshold of ice planting temperature

[0102] Using the DSC analysis software Pyris Manager to calculate the melting temperature of different concentrations of Me2SO4 solutions, the results are shown in the attached figure 2 As shown, the melting temperatures of different concentrations of Me2SO are shown here, where the melting temperatures of 0% Me2SO, 3% Me2SO, 5% Me2SO, and 10% Me2SO are -3.89±0.2°C, -6.08±0.49°C, -7.43±0.81°C, - 9.73±0.39°C. When the solution temperature is higher than the melting temperature, the solution will not freeze, only when the solution temperature is lower than the melting temperature, the solution has the possibility of freezing. According to the melting temperature and ensuring a certain margin, the temperature range of ultrasonic ice planting can be determined as follows: the upper threshold of the ice planting temperature of 0% Me2SO solution is about -4°C, ...

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Abstract

The invention relates to an ultrasonic plant freezing and storing device, in particular to an ultrasonic plant freezing and storing device for hepatocytes and a freezing and storing method of the ultrasonic plant freezing and storing device. The ultrasonic plant-freezing storage device comprises an ultrasonic container, a temperature detection system, an ultrasonic induced nucleation system and a cooling system, the liver cell ultrasonic plant-freezing preservation method comprises the following steps: culturing liver cells, determining an upper threshold of a plant-freezing temperature, selecting a temperature-controlled phase-change liquid raw material, and carrying out liver cell ultrasonic plant-freezing preservation. Compared with a conventional hepatocyte cryopreservation mode, the ultrasonic plant-freezing cryopreservation device and the ultrasonic plant-freezing cryopreservation method have the advantages that the hepatocyte cryopreservation effect is remarkably improved, the use amount of the cryoprotectant is small, the ultrasonic plant ice and the cryopreservation protectant have a synergistic effect, the recovery survival rate of the cryopreserved hepatocytes is higher than 90%, and no significant difference exists between the cryopreserved hepatocytes and fresh groups; and an unexpected technical effect is achieved.

Description

technical field [0001] The present invention relates to an ultrasonic implant freezing device, in particular to an ultrasonic implant freezing device for liver cells and a freezing method thereof. Background technique [0002] Cell cryopreservation is a key technology used in modern regenerative medicine, organ transplantation and assisted reproduction. Cell therapy is currently recognized as an effective means of fighting cancer and other intractable diseases, and has attracted widespread attention from all parties. The cryopreservation of cells is generally done by freezing the cells to a lower subzero temperature (usually -196°C). The cells are at low temperatures and have no vital activities, so they can be stored for a long time. However, severe cell damage may occur during the cooling process. damage. In order to reduce freezing damage, the commonly used cell freezing scheme is to add a certain concentration of cryoprotectant to the cell solution, and then freeze by s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 李维杰申敬宋立勇刘宝林漆琴
Owner UNIV OF SHANGHAI FOR SCI & TECH
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