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Cell gel preparation for treating articular cartilage injury and use thereof, and used gel solution for maintaining activity of cryopreserved cells

A technology of cell activity and gel solution, which is applied in the fields of biotechnology and biomedicine, can solve the problems of increasing the risk of cell contamination, and achieve the effects of good biocompatibility, high survival rate, and easy preparation and operation

Inactive Publication Date: 2018-10-19
TIANJIN AMCELLGENE ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method increases the risk of cell contamination and requires strict procedures

Method used

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  • Cell gel preparation for treating articular cartilage injury and use thereof, and used gel solution for maintaining activity of cryopreserved cells
  • Cell gel preparation for treating articular cartilage injury and use thereof, and used gel solution for maintaining activity of cryopreserved cells
  • Cell gel preparation for treating articular cartilage injury and use thereof, and used gel solution for maintaining activity of cryopreserved cells

Examples

Experimental program
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Effect test

preparation example Construction

[0038] The preparation method of the gel preparation containing mesenchymal stem cells and / or chondrocytes, the specific steps are as follows:

[0039] ① Preparation of gel solution:

[0040] According to the ratio provided above, the injection-grade or pharmaceutical-grade raw materials, glycerol, human serum albumin, DMSO, chondroitin sulfate, dissolved in compound electrolyte injection and / or amino acid injection, and normal saline injection are replenished, After fully stirring and mixing, set aside.

[0041] ②Acquired cells:

[0042] Select the required cells from the cell bank, use DMEM medium containing 10% fetal bovine serum as the complete medium, and store the cells at 37°C, 5% CO 2When the cells grow to about 85%, the cells are washed with phosphate buffer, digested with 0.25% trypsin, and then the digestion is stopped with complete medium, and the obtained cell suspension is subjected to 1500 × g for 5 min. Centrifuge, discard the supernatant, then resuspend the...

Embodiment 1

[0045] Example 1: Viability detection of umbilical cord mesenchymal stem cells in sodium hyaluronate / collagen / chondroitin sulfate (1:1:0.1) gel

[0046] Combine sterile sodium hyaluronate (10mg), collagen (10mg) and chondroitin sulfate (1mg) into a blended powder, dissolve it in 8.49ml of phosphate solution, swell at room temperature for 24 hours, then add 1.5ml of glycerin , 50mg of human serum albumin, 10ul DMSO, fully stirred and mixed to prepare a polymer material gel solution. After standing still for use, take 900ul gel solution and add 100ul containing 5×10 5 / cells The compound electrolyte resuspension of umbilical cord mesenchymal stem cells, mix well, and make sodium hyaluronate / chondroitin sulfate / collagen gel preparation. They were directly frozen and stored in a -80°C refrigerator. After 0, 7, and 15 days, 3 tubes were taken from each group for cell viability detection. It can be seen from the experimental results that the gel preparation has good cytocompatibil...

Embodiment 2

[0047] Example 2: Viability detection of umbilical cord mesenchymal stem cells in sodium hyaluronate / collagen / chondroitin sulfate gel (48:2:1)

[0048] Combine sterile sodium hyaluronate (480mg), collagen (20mg) and chondroitin sulfate (10mg) into blended powder, dissolve in 9.4ml amino acid injection, swell at room temperature for 24 hours, then add 300ul glycerol, 5mg of human serum albumin, 300ul DMSO, fully stirred and mixed, prepared polymer material gel solution. After standing still for use, take 900ul gel solution and add 100ul containing 5×10 5 / cells The compound electrolyte resuspension of umbilical cord mesenchymal stem cells, mix well, and make sodium hyaluronate / collagen / chondroitin sulfate gel preparation. They were directly frozen and stored in a -80°C refrigerator. After 0, 7, and 15 days, 3 tubes were taken from each group for cell viability detection. It can be seen from the experimental results that the gel preparation has good cytocompatibility, that is,...

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Abstract

The invention discloses a cell gel preparation for treating articular cartilage injury and a use thereof, and a used gel solution for maintaining the activity of cryopreserved cells. The formula of the cell gel preparation comprises, according to the volume percentage, 50% to 95%, of the gel solution and 5% to 50% of a compound electrolyte cell resuspension, and per milliliter of the cell gel preparation contains 0.5*10<5>-2*10<7> cells. The gel solution includes the following components by mass and volume: 1 to 50 mg of a high molecular material stabilizer, 0.1 to 1 mg of chondroitin sulfate,5 to 50 mg of human serum albumin, 30 to 150 [mu]l of glycerol, 1 to 30 [mu]l of DMSO, and 700 to 900 [mu]l of a compound electrolyte injection liquid or amino acid injection liquid or normal salineinjection liquid. The gel preparation has good biocompatibility and can be directly stored at low temperature for a long time and maintain high cell viability, and does not require traditional liquidnitrogen cryopreservation; the cell gel preparation after resuscitation can maintain cell biological characteristics and functions and high survival rate in a joint cavity, and can be used in treatment of articular cartilage injury.

Description

technical field [0001] The invention relates to the fields of biotechnology and biomedicine, in particular to a cell gel preparation for treating articular cartilage damage, its application, and a gel solution for maintaining the activity of frozen cells. Background technique [0002] Articular cartilage defects are very common in clinical practice in my country. According to statistics, there are nearly 10 million new patients with cartilage injuries in my country every year. Since the surface tissue of articular cartilage has no nerves, no blood vessels, and no lymphatic drainage, it is difficult for damaged cartilage tissue to repair itself. At present, the clinical methods for repairing articular cartilage damage include: joint fusion, joint debridement, arthroplasty, artificial joint replacement, autologous or allogeneic cartilage transplantation, matrix-induced autologous chondrocyte transplantation, and gene therapy and stem cells. therapy etc. Although this method ...

Claims

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Application Information

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IPC IPC(8): A61K9/06A61K35/28A61K35/32A61K47/36A61K47/42A61K47/20A61K47/10A61P19/02A61P19/00A01N1/02
CPCA01N1/0226A01N1/0231A61K9/06A61K35/28A61K35/32A61K47/10A61K47/20A61K47/36A61K47/42A61P19/00A61P19/02A61K2300/00
Inventor 韩之波喻昊贾红红韩忠朝
Owner TIANJIN AMCELLGENE ENG
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