A kind of tissue preservation solution and its application

A technology for preserving liquid and tissue, applied in the field of biomedicine, can solve the problem of inability to retain cell activity and source tissue for a long time, and achieve the effect of maintaining cell development characteristics, high cell viability, and improving success rate.

A technology for preserving liquid and tissue, applied in the field of biomedicine, can solve the problem of inability to retain cell activity and source tissue for a long time, and achieve the effect of maintaining cell development characteristics, high cell viability, and improving success rate.

CN112841168BActive Publication Date: 2021-12-10ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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  • A kind of tissue preservation solution and its application
  • A kind of tissue preservation solution and its application
  • A kind of tissue preservation solution and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] This example provides a method for preserving and culturing mouse liver tissue. The tissue preservation solution involved includes Advanced DMEM / F12 medium and added factors; the added factors include components at the following final concentrations: N-acetylcysteine, 5 μM ; Indole-3-lactic acid: 100μM; Y-27632 100μM; GDF11100ng / ml; penicillin-streptomycin mixture, 5×; Preservation and cultivation method specifically comprises the following steps:

[0033] (1) After the mice were killed, soak them in normal saline for 10 minutes, soak them in alcohol for 30 seconds, take the liver after dissection, and cut it into about 2mm with surgical scissors 3 Place these small pieces in the aforementioned tissue preservation solution and store them at 4°C for 12 hours;

[0034] (2) Then add 10ml collagenase to the tissue preservation solution in step (1) to resuspend, transfer to 37°C, 220rpm shaker for digestion for 20min, filter the cells with a 100um cell mesh, add 10ml DMEM / F...

Embodiment 2

[0036] This embodiment provides a method for preserving and culturing mouse intestinal cancer tissue, the tissue preservation solution involved includes Advanced DMEM / F12 medium and added factors; the added factors include the following components at final concentrations: N-acetylcysteine, 2μM; indole-3-lactic acid: 500μM; Y-27632 50μM; penicillin streptomycin mixed solution, 3×; Primocin, 2mg / ml; EGF, 50ng / ml; CHIR99021, 5μM; Mix it evenly into the base medium. Preservation and cultivation method specifically comprises the following steps:

[0037] (1) One PDX mouse model of intestinal cancer was taken, sacrificed and dissected to obtain tumor tissue. Surgical scissors shredded to 2mm 3 Place these small pieces in the aforementioned tissue preservation solution and store them at 4°C for 48 hours;

[0038] (2) Isolate tumor cells by the slide method, collect tumor cells and resuspend in DMEM / F12 medium, count the cells, and calculate the cell yield, then inoculate the cells...

Embodiment 3

[0040]This embodiment provides a method for preserving and culturing mouse lung tissue. The tissue preservation solution involved includes Advanced DMEM / F12 medium and added factors; the added factors include components at the following final concentrations: N-acetylcysteine, 0.2 μM; indole-3-lactic acid: 1000μM; Y-27632 10μM; GDF1110ng / ml; penicillin-streptomycin mixture, 1×; . Preservation and cultivation method specifically comprises the following steps:

[0041] (1) After the mice were killed, they were soaked in normal saline for 10 minutes, soaked in alcohol for 30 seconds, and the lungs were taken after dissection, and the bronchi were stripped as much as possible, and the lung parenchyma was harvested, and cut into about 1mm with surgical scissors 3 Place these small pieces in the aforementioned tissue preservation solution and store them at 4°C for 12 hours;

[0042] (2) Then add 10ml collagenase to the tissue preservation solution in step (1) to resuspend, transfer...

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Abstract

The invention provides a tissue preservation solution and its application. The tissue preservation solution includes basal culture solution and added factors; the added factors include at least the following final concentration components: N-acetylcysteine, 0.2-5 μM; indole-3-lactic acid: 100-1000 μM; Y-27632 10- 100μM; GDF11 10-100ng / ml; Penicillin-Streptomycin Mixture, 1-5×; Primocin, 0.5-5mg / ml. The tissue preservation solution is used in the preservation of isolated tissues before primary culture. The preservation method includes: completely immersing the tissue in the above-mentioned tissue preservation solution after being isolated, and preserving it at 2-8°C. The tissue preservation solution of the present invention has an effective storage period of more than 12 hours for normal tissues after isolation, and an effective storage period of tumor tissues and their cells for more than 48 hours after isolation, and tissues preserved for a long time can still maintain a high Cell viability and maintain cell development characteristics, significantly improve the success rate of primary cell culture. In addition, the tissue preservation solution of the present invention is of great significance to primary culture specimens that need to be preserved for a long time, especially precious clinical specimens.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a tissue preservation solution and its application. Background technique [0002] Primary culture is an important research method, including adherent culture, suspension culture, 3D culture and other methods. However, the success rate of primary culture has a great correlation with the activity of source tissue samples. However, due to the influence of various factors after the tissue is isolated, the cell viability decreases or even dies, which makes it impossible to obtain a sufficient amount of active cells during the pretreatment process of the primary culture, which leads to the failure of the culture. For example, the loss of contact with other cells in vivo or extracellular mechanisms induces the regulation of related apoptotic mechanisms after cells are isolated, resulting in anoikis. In addition, after organoids are isolated from the body, due to the lo...

Claims

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Application Information

Patent Timeline
10 Dec 2021
Publication
CN112841168B
IPC
A01N1/02
CPC
A01N1/021; A01N1/0226
Inventors
陈泽新; 朱宇