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In-vitro tissue preserving fluid and preserving method and application thereof

A preservation solution, in vitro technology, applied in the preservation, application, animal husbandry, etc. of human or animal body, can solve the problems of sample freezing failure, low tissue activity, low modeling success rate, etc. The effect of reducing repeated sampling and improving the success rate of modeling

Inactive Publication Date: 2020-04-28
南京普恩瑞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the preservation of the activity of isolated tissues by common preservation solutions needs to be improved, which will lead to low tissue activity, low modeling success rate, and sample cryopreservation failure.

Method used

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  • In-vitro tissue preserving fluid and preserving method and application thereof
  • In-vitro tissue preserving fluid and preserving method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: Preparation of isolated tissue preservation solution

[0023] Preparation of improved L-15 medium: based on L-15 medium, configure 10mM PBS buffer solution, the configuration method is the prior art; add galacto-oligosaccharides, SOD, vitamin E, glutathione, hydrocortisone Pine and IL-2, configured into the ratio of each component is as follows: 8% fetal bovine serum, 320mg / L L-glutamine, 780mg / L galactose, 35mg / L galactooligosaccharides, 400U / ml SOD , 42μg / L vitamin E, 400mg / L sodium pyruvate, 80mg / L glutathione, 1.1μg / L hydrocortisone and 700U / ml IL-2, the configuration method is the prior art.

[0024] Preservation solution: Dissolve 100 μL of penicillin-streptomycin double antibody in 1 tube of preservation solution, preparation of each tube of isolated tissue preservation solution: 100 μL of double-antibody (penicillin-streptomycin) + 3mL of fetal bovine serum + 6.9mL of Improve the L-15 medium, see Table 1 below for details.

[0025] Formulation of...

Embodiment 2

[0028] Embodiment 2: Preparation of isolated tissue preservation solution

[0029] Preparation of improved L-15 medium: based on L-15 medium, configure 10mM PBS buffer solution, the configuration method is the prior art; add galacto-oligosaccharides, SOD, vitamin E, glutathione, hydrocortisone Pine and IL-2, configured into the ratio of each component as follows: 8% fetal bovine serum, 330mg / L L-glutamine, 800mg / L galactose, 40mg / L galactooligosaccharides, 500U / ml SOD , 40μg / L vitamin E, 420mg / L sodium pyruvate, 90mg / L glutathione, 1.2μg / L hydrocortisone and 800U / ml IL-2, the configuration method is the prior art.

[0030] Preservation solution: Dissolve 200 μL of penicillin-streptomycin double antibody in 1 tube of preservation solution, preparation of each tube of isolated tissue preservation solution: 100 μL of double-antibody (penicillin-streptomycin) + 3mL of fetal bovine serum + 6.8mL of For improved L-15 medium, see Table 2 below for details.

[0031] Formulation of t...

Embodiment 3

[0033] Example 3: Preparation of isolated human gastrointestinal tumor tissue

[0034] A method for preparing isolated human gastrointestinal tumor tissue, the steps comprising:

[0035] (1) Material collection: take patients' gastrointestinal tumor tissue through surgery, puncture, and endoscopy, and place it in the preservation solution prepared in Example 2. It is necessary to completely soak the tissue sample in the preservation solution to avoid excessive tissue volume and Part of the tissue is not immersed in the preservation solution or the tissue is separated from the preservation solution due to factors such as bumps during transportation;

[0036] (2) Tissue transportation: transport the tissue back to the laboratory in a low-temperature environment (place ice packs in an insulated bucket) for no more than 24 hours;

[0037] (3) Tissue processing: put the tumor tissue in a petri dish, add L-15, and place it on ice for later use; observe the tumor tissue under a diss...

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Abstract

The invention discloses an in-vitro tissue preserving fluid as well as a preserving method and application thereof. The in-vitro tissue preserving fluid includes a penicillin-streptomycin solution, fetal calf serum and an improved L-15 culture medium. The obtained in-vitro tissue preserving fluid can provide nutrition for tumor tissues and has an antibacterial effect, the activity of the tumor tissues can be improved, the modeling success rate of clinical patient-derived tumor xenograft is further improved, in addition, tumor tissue samples can be preserved, a sample database can be established, secondary experiment needs are facilitated, and repeated material taking is reduced.

Description

technical field [0001] The invention relates to the technical field of biological reagents, in particular to an isolated tissue preservation solution and its preservation method and application. Background technique [0002] Tissue inoculation, tissue cryopreservation and resuscitation have been widely used in many fields such as medical treatment, biology, eugenics, etc. However, for some difficult-to-obtain tissues, due to experimental work, repeated experiments, repeated cultivation, or direct inoculation with tissue blocks are required into the animal body. [0003] At present, the preservation of the activity of isolated tissues by ordinary preservation solutions needs to be improved, which will lead to low tissue activity, low success rate of modeling and failure of sample cryopreservation. [0004] The information disclosed in this Background section is only for enhancing the understanding of the general background of the present invention and should not be taken as ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0215A01N1/0226
Inventor 朱燕萍王志强郝艳鹏李晓宇叶琳璐
Owner 南京普恩瑞生物科技有限公司
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