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Stem cell cryopreservation liquid and stem cell cryopreservation method

A cryopreservation method and stem cell technology, applied in the field of stem cell cryopreservation and stem cell cryopreservation, can solve the problems that stem cells are easily damaged by ice crystals, cells are easily aged, and have low efficiency, so as to achieve unaffected activity and cell proliferation ability, avoid The effect of excessive dehydration and shrinkage of cells and improving the effect of cryopreservation

Inactive Publication Date: 2022-04-12
GUANGDONG XIANKANGDA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the existing cryopreservation methods of stem cells, only the cryopreservation of stem cells and tumor cells has been solved, and the method of stem cell cryopreservation has not been well solved.
Due to the cell structure and cell characteristics of stem cells, when using the previous cell cryopreservation solution and cryopreservation method to freeze stem cells, the stem cells are easily damaged by ice crystals. After recovery, the cell viability is low, the number of cell proliferation is insufficient, the cells are easy to age, and the efficiency is low. The quality of cryopreservation is not high, the activity of cells after recovery is poor, and the survival rate is not high.

Method used

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  • Stem cell cryopreservation liquid and stem cell cryopreservation method
  • Stem cell cryopreservation liquid and stem cell cryopreservation method
  • Stem cell cryopreservation liquid and stem cell cryopreservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] ①. In the biological safety cabinet or ultra-clean workbench, take out the umbilical cord, place it together with the umbilical cord clamp in a 10ml centrifuge tube or a 10cm petri dish, soak in 75% alcohol for 2 minutes;

[0082] ② After treatment with alcohol, quickly move the umbilical cord into a container filled with PBS or saline, soak and wash twice, 2 minutes each time;

[0083] ③. Cut off the umbilical cord clamp, cut the umbilical cord into small sections, each 2-3cm, cut the umbilical cord from the inside of the umbilical vein with scissors, and remove the umbilical vein, umbilical artery and umbilical cord capsule with toothed forceps to prevent the mixing of miscellaneous cells;

[0084] ④. Shred the umbilical cord tissue with scissors, digest with 0.1% type II collagenase (diluted with DMEM / F12) for 2 hours, and digest on a shaker at 37°C;

[0085] ⑤. Rinse with PBS or saline several times to remove the collagenase attached to the surface of the tissue as ...

Embodiment 2

[0093] (1) Configuration of cryopreservation solution

[0094] Commercially available serum-free GT-T551H3 medium, which contains α-MEM basal medium; add human albumin, sulfosalicylic acid, acetamide and dimethyl sulfoxide to the medium, and mix well Wherein, in the culture medium, the final volume percentage content of human albumin, sulfosalicylic acid, acetamide and dimethyl sulfoxide is shown in Table 1.

[0095] Table 1 Configuration list of frozen storage solution components

[0096] component name contains components Volume percentage, % Serum-free GT-T551H3 medium Contains 55% α-MEM basal medium 62 Add human serum albumin at a concentration of 25 g / l - 18 sulfosalicylic acid - 2 Acetamide - 8 Dimethyl sulfoxide - 10

[0097] (2) Stem cell cryopreservation

[0098] After the umbilical cord stem cells in Example 1 were resuspended and mixed evenly, the cells were counted, the required cell suspension was divided int...

Embodiment 3

[0105] Commercially available serum-free GT-T551H3 medium, which contains α-MEM basal medium; add human albumin, sulfosalicylic acid, acetamide and dimethyl sulfoxide to the medium, and mix well ; Wherein, in the culture medium, the final volume percentage contents of human albumin, sulfosalicylic acid, acetamide and dimethyl sulfoxide are shown in Table 2.

[0106] Table 2 Component Configuration Table of Freezing Solution

[0107] component name contains components Volume percentage, % Serum-free GT-T551H3 medium Contains 50% α-MEM basal medium 53 Add human serum albumin at a concentration of 25 g / l - 20 sulfosalicylic acid - 5 Acetamide - 10 Dimethyl sulfoxide - 12

[0108] (2) Stem cell cryopreservation

[0109] After the umbilical cord stem cells in Example 1 were resuspended and mixed evenly, the cells were counted, the required cell suspension was divided into centrifuge tubes, the supernatant was removed by centrif...

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Abstract

The invention provides a stem cell cryopreservation liquid and a stem cell cryopreservation method, and the stem cell cryopreservation liquid comprises the following components by volume percentage: a serum-free GT-T551H3 culture medium containing 50%-60% of an alpha-MEM basic culture medium; 15%-20% of human serum albumin with a concentration of 15-35 g / l; 1% to 5% of sulfosalicylic acid; 5% to 10% of acetamide; 8%-12% of dimethyl sulfoxide; wherein the volume ratio of sulfosalicylic acid to acetamide is (1: 1)-(1: 10). The stem cell cryopreservation liquid provided by the invention has the advantages; the cell cryopreservation liquid does not contain animal serum, so that the risk of introducing pollution and allergens can be avoided, and the cell cryopreservation liquid has higher clinical safety compared with a conventional cell cryopreservation liquid; and the sulfosalicylic acid and acetamide components can prevent the cells from being damaged due to formation of ice crystals in the cryopreservation process.

Description

technical field [0001] The invention relates to the technical field of cell cryopreservation, and more specifically, to a stem cell cryopreservation solution and a stem cell cryopreservation method. Background technique [0002] Stem cells are a kind of multifunctional cells with multi-directional differentiation potential and self-renewal ability. They are the most primitive cells at the top of the origin of cell lines and can differentiate into cells of a specific tissue type in vivo. In adult organs, stem cells can repair tissue by continually dividing. [0003] The damage and failure of tissues and organs has always been a major problem facing human health. It has always been a human goal to perfectly repair or replace the damage to tissues, organs or limbs caused by diseases, war injuries, accidents or genetic factors. A dream is also a medical peak that is difficult to overcome. Current treatment options are difficult to completely repair damaged tissues and organs o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226A01N1/0284
Inventor 李勇刘家飞薛卫巍谢海涛
Owner GUANGDONG XIANKANGDA BIOTECH CO LTD
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