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Tumor tissue cryopreservation, resuscitation kit and processing method thereof

A tumor tissue and processing method technology, applied in the field of tumor tissue processing, can solve the problems of easy loss of tumor original mutation, short shelf life, harsh storage conditions, etc., and achieve the effect of improving the survival rate of resuscitation, easy operation, and simplified operation

Active Publication Date: 2018-08-03
SHANGHAI CRYOWISE MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the variable composition of serum and the poor stability of the cryopreservation solution, the shelf life of this tissue cryopreservation solution is short and the storage conditions are relatively harsh.
[0005] When using the existing tissue cryopreservation solution to freeze tumor tissue, only a very small number of tumor tissues can be revived or passaged in the form of tumor cells, let alone proliferate in the form of primary tumors, and the original tumor mutations are easily lost. Only data information at the molecular level can be obtained
Therefore, the application range of the existing tissue cryopreservation solution is narrow, which reduces the application value and scientific transformation ability of tumor tissue samples

Method used

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  • Tumor tissue cryopreservation, resuscitation kit and processing method thereof
  • Tumor tissue cryopreservation, resuscitation kit and processing method thereof
  • Tumor tissue cryopreservation, resuscitation kit and processing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: Preparation of Tumor Tissue Cryopreservation Kit

[0057] Prepare the vitrification solution required for tumor tissue cryopreservation, the specific formula components are as follows:

[0058] Vitrification solution 1 (V1), composed of DMEM80V / V%, DMSO 10V / V%, EG10V / V%, BSA3w / v%, sucrose 1w / v%, methylcellulose 4000CP0.05w / v%, hydroxyethyl Starch 0.25w / v%, glucose 25w / v%.

[0059] Vitrification solution 2 (V2), composed of DMEM70V / V%, DMSO18V / V%, EG12V / V%, BSA 3w / v%, sucrose 20w / v%, methylcellulose 4000CP0.05w / v%, PVP0.25w / v%, glucose 30w / v% composition.

[0060] Among them, the Chinese name of each reagent and the manufacturer:

[0061] DMEM: High Glucose Modified Duchenne Eagle's Medium; Sigma-Aldrich

[0062] DMSO: Dimethylsulfoxide; Sigma-Aldrich

[0063] BSA: bovine serum albumin; Sigma-Aldrich

[0064] EG: Ethylene glycol; Sigma-Aldrich

[0065] Sucrose: Sigma-Aldrich

[0066] Methylcellulose 4000CP: Sigma-Aldrich

[0067] Hydroxyethyl starch:...

Embodiment 2

[0070] Example 2: Cryopreservation of tumor tissue

[0071] The prepared cryopreservation reagents were filtered, aliquoted and stored in a -4°C refrigerator, and stored in a -20°C refrigerator for long-term storage.

[0072] Cryopreservation of tumor tissue, the specific steps are as follows:

[0073] a. When the tumor tissue is transported to the treatment room on ice with transport fluid, wash it three times with normal saline, trim and remove blood vessels, capsules, necrotic tissue, etc., use the tumor tissue processing mold to slice into tissue pieces with a size of 1x10x10mm, and use Wash again with normal saline;

[0074] Among them, the tumor tissue processing mold such as Figure 5 As shown, the tumor tissue processing mold includes: a base 1, the upper surface of the base 1 is provided with a depression 2, and vertically downward from the upper surface of the base 1, 12 equidistant distributions are arranged along the lateral direction, and the depth is lower than...

Embodiment 3

[0080] Example 3: Preparation of Tumor Tissue Recovery Kit

[0081] Prepare the resuscitation solution required for tumor tissue resuscitation, the specific formula components are as follows:

[0082] Resuscitation solution 1 (T1), composed of DMEM65V / V%, 1XPBS35V / V%, BSA2w / v%, sucrose 40w / v%, glucose 25w / v%;

[0083] Resuscitation solution 2 (T2), composed of DMEM75V / V%, 1XPBS25V / V%, BSA2w / v%, sucrose 20w / v%, glucose 25w / v%;

[0084] Resuscitation solution 3 (T3) consists of DMEM95V / V%, 1XPBS5V / V%, BSA2w / v%, glucose 15w / v%.

[0085] Among them, the Chinese name of each reagent and the manufacturer:

[0086] DMEM: High Glucose Modified Duchenne Eagle's Medium; Sigma-Aldrich

[0087] 1XPBS: 0.01M Phosphate Buffered Saline; Sigma-Aldrich

[0088] BSA: bovine serum albumin; Sigma-Aldrich

[0089] Sucrose: Sigma-Aldrich

[0090] Glucose: Sigma-Aldrich

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Abstract

The invention relates to the technical field of treatment of living tumor tissue, and provides a kit for cryopreservation and resuscitation of living tumor tissue and a treatment method thereof. The cryopreservation and resuscitation kit of the present invention includes tissue vitrification fluid and resuscitation fluid, and the components contained are completely clear, stable and controllable, no serum substances are added, the shelf life is long, the stability between batches is good, and it can be directly applied It can be used for cryopreservation and recovery of living tissue without dilution or self-preparation, which simplifies the operation. The invention can make the tumor tissue cryopreserved and resuscitated to maintain a high survival rate, and preserve the original characteristics of the tumor tissue. It broadens the application scope of tumor tissue samples and better serves clinical research.

Description

technical field [0001] The invention relates to the technical field of tumor tissue processing, in particular to a tumor tissue cryopreservation and resuscitation kit and a processing method thereof. Background technique [0002] Living tissue, especially living tumor tissue, has important scientific research and clinical application values, and living tissue preservation technology is the basis for realizing these values. The commonly used preservation methods of living tumor tissues are mainly to establish PDX tumor model preservation and cryopreservation. Establishing a PDX tumor model for preservation is time-consuming, labor-intensive, cumbersome, and expensive. After multiple passages, the tumor tissue is prone to mutation, the heterogeneity of the tumor tissue is lost, and the value of preservation is lost. [0003] The cryopreservation of living tumor tissue is to store the tissue in a low-temperature environment, so that the tissue can temporarily escape from the g...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09A01N1/02
CPCA01N1/0231A01N1/0284C12N5/0693C12N2500/34C12N2500/30C12N2500/05A01N1/0268A01N1/0221C12N5/0018G01N1/06C12N2533/78C12N2523/00A01N1/0242A01N1/0278
Inventor 吴红平曾敏周徐金晓芳杨秋蕊
Owner SHANGHAI CRYOWISE MEDICAL TECH CO LTD
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