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Vitrified cryopreservation method for cartilage

A technology of vitrification and cartilage, applied in the field of medical pathology, can solve imperfections and other problems, achieve the effect of reducing toxicity damage and prolonging storage time

Inactive Publication Date: 2017-05-10
WEIFANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the application of vitrification in cartilage preservation is not perfect, and there are still many areas that need to be improved and improved

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A method for vitrifying and freezing cartilage, comprising the steps of:

[0024] 1) Hanks balanced salt solution per 1000ml of vitrification solution also includes: DMSO20.5mmol, acetamide 15.5mmol, propylene glycol 10mmol, galactose 3g, penicillin 90U and polyethylene glycol 6mmol, with 2mol / L sodium hydroxide Adjust the pH to 7.4. Adjust the pH to 7.4 with 2mol / L sodium hydroxide.

[0025] 2) Cell culture The collected cartilage was fully rinsed with sterilized PBS, and cell culture and cell passage were performed to obtain a cell suspension.

[0026] 3) Cryopreservation Pre-cool the cell suspension and vitrification solution to 4°C for 2 hours respectively, slowly add the vitrification solution to the cell suspension to obtain the cell suspension, the dropping process is: 0.3ml / min for the first 3 minutes Add dropwise, add dropwise at 0.6ml / min for the last 5 minutes, and add dropwise at 0.75ml / min for the remaining cryopreserved solution. Divide the frozen cell ...

Embodiment 2

[0030] A method for vitrifying and freezing cartilage, comprising the steps of:

[0031] 1) Prepare vitrification solution, add 20mmol of DMSO, 16mmol of acetamide, 8mmol of propylene glycol, 4g of galactose, 5mmol of polyethylene glycol and 70U of penicillin to each 1000ml of Hanks balanced salt solution, adjust the pH to 7.4.

[0032] 2) Cell culture The collected cartilage was fully rinsed with sterilized PBS, and cell culture and cell passage were performed to obtain a cell suspension.

[0033] 3) Cryopreservation Pre-cool the cell suspension and vitrification solution to 4°C for 2 hours respectively, slowly add the vitrification solution to the cell suspension to obtain the cell suspension, the dropping process is: 0.3ml / min for the first 3 minutes Add dropwise, add dropwise at 0.6ml / min for the last 5 minutes, and add dropwise at 0.75ml / min for the remaining cryopreserved solution. Divide the frozen cell suspension into cryovials, seal both ends of the cryovials with f...

Embodiment 3

[0036] A method for vitrifying and freezing cartilage, comprising the steps of:

[0037] 1) Prepare vitrification liquid and add 18mmol of DMSO, 14mmol of acetamide, 8mmol of propylene glycol, 2g of galactose, 5mmol of polyethylene glycol and 75U of penicillin to each 1000ml of Hanks balanced salt solution, adjust the pH to 7.4 with 2mol / L sodium hydroxide .

[0038] 2) Cell culture The collected cartilage was fully rinsed with sterilized PBS, and cell culture and cell passage were performed to obtain a cell suspension.

[0039] 3) Cryopreservation Pre-cool the cell suspension and vitrification solution to 4°C for 2 hours respectively, slowly add the vitrification solution to the cell suspension to obtain the cell suspension, the dropping process is: 0.3ml / min for the first 3 minutes Add dropwise, add dropwise at 0.6ml / min for the last 5 minutes, and add dropwise at 0.75ml / min for the remaining cryopreserved solution. Divide the frozen cell suspension into cryovials, seal bo...

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PUM

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Abstract

The invention discloses a vitrified cryopreservation method for cartilage. The method comprises the following steps that 1, a vitrification solution is prepared, wherein DMSO, acetamide, propylene glycol, galactose, penicillin and polyethylene glycol are added into a Hanks balanced salt solution, and the pH value is regulated to be 7.3-7.5 with sodium hydroxide with the concentration of 2 mol / L; 2, cell culture is conducted to obtain cell suspension; 3, cryopreservation is conducted, the frozen cell suspension is subpackaged in freezing tubes, and the freezing tubes are quickly put into liquid helium to be preserved. According to the cryopreservation method, toxic damage of the vitrification solution is reduced, the survival rate of cartilage cells is increased, and the bioactivity of the cartilage cells is improved.

Description

technical field [0001] The invention relates to the field of medical pathology, in particular to a method for vitrified and frozen cartilage preservation. Background technique [0002] In recent years, with the improvement of people's living standards in our country and the comprehensive popularization of sports, the number of patients with articular cartilage injuries has increased significantly. Cartilage transplantation is one of the very promising repair methods for the treatment of cartilage lesions. Because the allogeneic cartilage graft material is relatively easy to obtain and can be prefabricated into any shape and size, it is convenient to make the normal functional articular cartilage into various shapes and implant it in the bone defect to restore the shape of the cartilage. [0003] In recent years, the technique of allogeneic osteochondral transplantation has attracted the attention of clinical scholars. In the process of articular cartilage transplantation, ...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 马晓君
Owner WEIFANG MEDICAL UNIV
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