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Tumor tissue/or cell cryopreservation and resuscitation kit and treatment method thereof

A technology of tumor tissue and processing method, which is applied in the field of gravity tissue cryopreservation, resuscitation kit and its processing, which can solve the problems of waste, loss of activity, and inability to preserve active tumor tissue cells with high activity, so as to achieve clear components and facilitate The effect of simple and clear operation and popular application and processing method

Active Publication Date: 2021-07-23
江苏安泰康健康科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Commonly used methods for preserving viable tumor tissues and / or cells, such as propylene glycol slow programmed freezing, direct injection of liquid nitrogen (ultra-fast freezing) and dimethyl sulfoxide vitrification will completely or partially preserve viable tumor tissues and / or cells. Loss of activity, resulting in a large amount of valuable clinically active tumor tissue and / or cells being wasted due to the inability to obtain long-term effective high-activity preservation

Method used

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  • Tumor tissue/or cell cryopreservation and resuscitation kit and treatment method thereof
  • Tumor tissue/or cell cryopreservation and resuscitation kit and treatment method thereof
  • Tumor tissue/or cell cryopreservation and resuscitation kit and treatment method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1, preparation of tumor tissue / or cell cryopreservation kit

[0060] Prepare the vitrification solution required for tumor tissue / or cell cryopreservation, the specific formula components are as follows:

[0061] Vitrification solution 1 includes: Gibco DMEM medium 80V / V%, dimethyl sulfoxide (DMSO) 10V / V%, ethylene glycol (EG) 10V / V%, bovine serum albumin (BSA) 2W / V% , sucrose 1W / V%, methylcellulose (4000CP) 0.05W / V%, hydroxyethyl starch 0.25W / V% and glucose 25W / V%.

[0062] Vitrification solution 2 includes: DMEM medium 70V / V%, dimethylsulfoxide (DMSO) 15V / V%, ethylene glycol (EG) 12V / V%, bovine serum albumin (BSA) 3W / V%, Sucrose 20W / V%, methylcellulose (4000CP) 0.1W / V%, polyvinylpyrrolidone (PVP) 0.25W / V%, and glucose 30W / V%.

[0063] Among them, the Chinese and English names and purchasing manufacturers of each reagent are listed in Table 1:

[0064] Table 1

[0065]

[0066]

Embodiment 2

[0067] Embodiment 2, cryopreservation processing of tumor tissue / or cell

[0068] Filter and aliquot the prepared frozen reagents and store in a -4°C refrigerator; if the prepared frozen reagents need to be stored for a long time, they should be stored in a -20°C refrigerator.

[0069] Cryopreservation of tumor tissue / or cells, the methods of cryopreservation include:

[0070] Step S1, the tumor tissue / or cell sample is washed twice with 1x phosphate buffered saline (English abbreviation: 1xPBS) to remove the capsule, blood vessels and mucinous necrotic tissue; the operation should be done gently to avoid tumor tissue / or Cell mass damage. In addition, when the tumor tissue / or cell sample is processed for blood vessels, capsule and necrotic tissue, the processed tumor tissue / or cell sample must be completely immersed in 1x phosphate buffer.

[0071] Step S2, using the tumor slicing mold to slice the tumor tissue / or cells, and wash again with 1x phosphate buffered saline. Whe...

Embodiment 3

[0080] Example 3, Preparation of tumor tissue / or cell recovery kit

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

[0082] The components of resuscitation solution 1 include: Earl’s balanced salt solution 65V / V%, 1x phosphate buffer 35V / V%, bovine serum albumin 1w / v%, sucrose 30w / v% and glucose 25w / v%;

[0083] The components of resuscitation solution 2 include: Earl's Balanced Salt Solution 75V / V%, 1x Phosphate Buffer Saline 25V / V%, Bovine Serum Albumin 1w / v%, Sucrose 15w / v% and Glucose 25w / v%; and

[0084]The components of resuscitation solution 3 include: Earl’s Balanced Salt Solution 95V / V%, 1x Phosphate Buffer Saline 5V / V%, Bovine Serum Albumin 1w / v%, Sucrose 10w / v% and Glucose 15w / v%.

[0085] Among them, the volume concentration gradient of Earl's balanced salt solution used in resuscitation fluid 1, resuscitation fluid 2 and resuscitation fluid 3 becomes larger, and durin...

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Abstract

The invention discloses a tumor tissue / or cell cryopreservation and resuscitation kit and a treatment method thereof.The tumor tissue / or cell cryopreservation and resuscitation kit comprises a cryopreservation kit body and a resuscitation kit body, the cryopreservation kit body comprises a vitrified liquid, the resuscitation kit body comprises a resuscitation liquid, and components contained in the vitrified liquid and the resuscitation liquid are completely clear, stable and controllable; the kit can be directly applied to cryopreservation and resuscitation of active tumor tissues / or cells, dilution or self-preparation is not needed, and the operation is simplified. Before and after the vitrification cryopreservation of the tumor tissues / cells, the biological activity and morphological characteristics of the tumor tissues / cells are not obviously changed, so that the vitrification cryopreservation and resuscitation solution system better maintains the microenvironment and tumor heterogeneity characteristics of the tumor tissues / cells; and in addition, the cryopreserved tumor tissues / or cells are subjected to orthotopic transplantation after being recovered, the tumor formation rate is high, and the viable tumor tissues / or cells can be effectively frozen.

Description

technical field [0001] The present invention relates to the technical field of tumor tissue / or cell processing, in particular to gravity tissue cryopreservation, recovery kit and processing method thereof. Background technique [0002] Vitrification is a new type of cryopreservation technology, which has potential application value for clinical live tumor tissue / or cell samples in scientific research and clinical fields. Commonly used methods for preserving viable tumor tissues and / or cells, such as propylene glycol slow programmed freezing, direct injection of liquid nitrogen (ultra-fast freezing) and dimethyl sulfoxide vitrification will completely or partially preserve viable tumor tissues and / or cells. Loss of activity, resulting in a large amount of valuable clinically active tumor tissue / or cells being wasted due to the inability to obtain long-term effective high-activity preservation. [0003] Vitrification solution was developed by Rau and Fahy as early as 1985, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02C12N5/09
CPCA01N1/0226A01N1/0221C12N5/0693C12N2500/34C12N2500/30
Inventor 孙晓娇刘星张红霞陆重益张文平
Owner 江苏安泰康健康科技有限公司
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