Serum-free cryopreservation liquid, and preparation method and preparation device thereof

A serum-free, cryopreserved liquid technology, applied in the preservation, application, animal husbandry and other directions of human or animal body, can solve the problems such as troublesome preparation, complex additive components, etc., and achieves simple preparation method, good application prospect, and survival rate. high effect

Inactive Publication Date: 2021-10-01
南京生航生物技术有限公司
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a serum-free cell cryopreservation solution and its preparation method and preparation device, aiming to solve the technical problems of complex ingredients and troublesome preparation in the cell cryopreservation solution sold in the prior art

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Serum-free cryopreservation liquid, and preparation method and preparation device thereof
  • Serum-free cryopreservation liquid, and preparation method and preparation device thereof
  • Serum-free cryopreservation liquid, and preparation method and preparation device thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0036] see figure 1 and figure 2 , the present invention provides a serum-free cell cryopreservation solution and its preparation method and preparation device, comprising the following components:

[0037] DMEM / F-12 basal medium, Nutridoma 0.2-1mg / ml, pyruvate 5-20mg / L, L-glutamine 3-6mg / ml, intracellular and extracellular cryoprotectant 0.1-0.41g / L; the cells The internal and external cryoprotectant is a mixed solution composed of trehalose 0.05g / L, polyvinylpyrrolidone 0.5mg / L and methylcellulose 0.4g / L.

[0038] In this embodiment, the cell cryopreservation solution improves the thawing survival rate of the cells cryopreserved by the cell cryopreservation solution, and can maintain good cell activity and physiological characteristics.

[0039] Cryopreservation of human umbilical cord mesenchymal stem cells: separate and culture human umbilical cord mesenchymal stem cells (MSCs) using the tissue block adherence method, spread them in a 10cm culture dish, add DMEM / F12 cul...

Embodiment 1

[0045]Example 1, see image 3 , the present invention also provides a preparation method using the above-mentioned serum-free cell cryopreservation solution, comprising the following steps:

[0046] S1: Take 1L DMEM / F-12 liquid basal medium;

[0047] S2: Weigh 5 mg of pyruvate, 3 mg of L-glutamine, 0.05 g of trehalose, 0.5 mg of polyvinylpyrrolidone, 0.2 g of methylcellulose, and 0.2 mg of Nutridoma, and dissolve them in 1 L of DMEM / F-12 liquid basal medium, Obtain the first mixed solution;

[0048] S3: adjusting the pH value of the first mixed solution to 7.0 to obtain a second mixed solution;

[0049] S4: After filtering the second mixed solution, a serum-free cell cryopreservation solution is obtained.

[0050] Among them, in step S1, DMEM / F-12 liquid basal medium is used, so that after the cells are recovered, there is no need to pour out the frozen storage solution before culturing. It can be directly added to the culture medium, and it can be passaged or used after it...

Embodiment 2

[0054] Example 2, see Figure 4 , the present invention also provides a preparation method using the above-mentioned serum-free cell cryopreservation solution, comprising the following steps:

[0055] S1: Take 1L DMEM / F-12 liquid basal medium;

[0056] S2: Weigh 12 mg of pyruvate, 4 mg of L-glutamine, 0.05 g of trehalose, 0.5 mg of polyvinylpyrrolidone, 0.2 g of methylcellulose, and 0.6 mg of Nutridoma, and dissolve them in 1L DMEM / F-12 liquid basal medium, Obtain the first mixed solution;

[0057] S3: adjusting the pH value of the first mixed solution to 7.2 to obtain a second mixed solution;

[0058] S4: After filtering the second mixed solution, a serum-free cell cryopreservation solution is obtained.

[0059] Among them, in step S1, DMEM / F-12 liquid basal medium is used, so that after the cells are recovered, there is no need to pour out the frozen storage solution before culturing. It can be directly added to the culture medium, and it can be passaged or used after it ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to the technical field of cell cryopreservation, and concretely relates to serum-free cell cryopreservation liquid, and a preparation method and a preparation device thereof. The cryopreservation liquid comprisesa DMEM/F-12 basic culture medium, Nutridoma, pyruvic acid, L-glutamine and 0.1-0.41 g/L of an intracellular and extracellular cryoprotectant. The cell cryopreservation liquid without serum or DMSO is determined in formula components and has very high biological safety and clinical application prospects, the recovery survival rate of cells cryopreserved with the cell cryopreservation liquid reaches 87% or above, good cell viability and physiological characteristics can be maintained, and the cryopreservation liquidis very suitable for the related research field of primary cells and stem cells.

Description

technical field [0001] The invention relates to the technical field of cell cryopreservation, in particular to a serum-free cell cryopreservation solution, a preparation method and a preparation device. Background technique [0002] Cell cryopreservation solution, as a solution that must be used for cell cryopreservation, is used to suspend cells that need to be frozen in the cryopreservation solution, supply the nutrients necessary for cell life and metabolism, and prevent or reduce the impact of frozen ice crystals on the cells. cell damage. The prior art generally uses a method of mixing culture medium, serum and dimethyl sulfoxide (DMSO) in a certain ratio to freeze and preserve cells. The amount of DMSO is generally 10%, but too low DMSO concentration will lead to poor freezing effect, while high concentration of DMSO will cause toxicity and damage the cell body; and the use of serum will increase the possibility of animal pathogen contamination, It cannot be directly...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0226A01N1/0221
Inventor 赵光锋李春慧李明扬王洋洋
Owner 南京生航生物技术有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap