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Stem cell preparation and preparation method and application hereof

A stem cell preparation, stem cell technology, applied in the field of stem cell preparation and its preparation, can solve the problems of periodontitis that cannot be cured, inconvenience for patients, easy recurrence, etc., achieve process optimization, improve quality of life, and low immunogenicity

Inactive Publication Date: 2017-10-17
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned methods all bring inconvenience and pain to the patient to a certain extent, and they cannot cure periodontitis, and it is easy to relapse or continue to aggravate

Method used

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  • Stem cell preparation and preparation method and application hereof
  • Stem cell preparation and preparation method and application hereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Isolation and culture of dental pulp mesenchymal stem cells

[0018] Collect the teeth, put the teeth in a 12-well plate after cleaning, put each tooth in only one well, wash twice with 3mL alcohol to remove periodontal ligament and tartar, and then wash with 3mL PBS containing 1% double antibody for 2 The next time, wrap the teeth with gauze, use a vise to clamp the teeth open, use a pointed pinch to clamp out the pulp tissue in the tooth cavity, put it in the EP tube, and then use small scissors to cut the tissue pieces into pieces.

[0019] Add 0.9mL type I collagenase with a volume percentage concentration of 0.3-0.5% and 0.6mL volume percentage concentration of 1% Dispase into a 15mL centrifuge tube, mix well, and transfer the pulp tissue to this centrifuge tube. tube, transferred to a constant temperature shaker for digestion at 37°C, 100-150rpm for 30-50min. Then add 2-4mL DMEM / F12 complete medium to stop the digestion, centrifuge at 1500rpm for 5min, ...

Embodiment 2

[0021] The third-generation dental pulp stem cells were washed twice with normal saline, digested with 0.25% trypsin, terminated with DMEM / F12 complete medium, transferred to a 50mL centrifuge tube and centrifuged at 1500rpm for 5min; resuspended cells in normal saline, 1500rpm Centrifuge for 5 minutes, remove the supernatant, and repeat this twice, then add physiological saline and human serum albumin, mix well, filter with a 70 μm filter, and collect the filtrate with a syringe to make a 10 mL injection preparation. Wherein, the cell density of the dental pulp mesenchymal stem cells in the injection preparation is 4×10 6 Individual / mL, the volume percentage concentration of human serum albumin is 5%.

Embodiment 3

[0023] Take the fourth-generation dental pulp stem cells and wash them twice with normal saline, digest them with 0.25% trypsin, stop the digestion with DMEM / F12 complete medium, transfer to a 50mL centrifuge tube and centrifuge at 1500rpm for 5min; resuspend the cells in normal saline, 1500rpm Centrifuge for 5 minutes, remove the supernatant, and repeat this twice, then add physiological saline and human serum albumin, mix well, filter with a 70 μm filter, and collect the filtrate with a syringe to make a 10 mL injection preparation. Wherein, the cell density of the dental pulp mesenchymal stem cells in the injection preparation is 5×10 6 Individual / mL, the volume percentage concentration of human serum albumin is 10%.

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Abstract

The invention belongs to the technical field of stem cells, and particularly relates to a stem cell preparation and a preparation method and application hereof. The stem cell preparation comprises dental pulp mesenchymal stem cells and human albumin. Components are reasonably matched and are synergistic, reparative treatment of periodontitis is promoted, and life quality of patients is improved. The dental pulp mesenchymal stem cells are adopted as seed cells, thereby being simple to collect, easy to obtain, low in immunogenicity and free of ethical issue and being of important significance for the patients with periodontitis. The preparation method is simple in step operation, optimized in process and suitable for large-scale production.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and in particular relates to a stem cell preparation and its preparation method and application. Background technique [0002] Periodontitis is a chronic inflammation that invades the gums and periodontal tissue. It is a destructive disease, and its main features are the formation of periodontal pockets, inflammation of the pocket wall, alveolar bone resorption and gradual loosening of teeth. main cause of loss. The disease is mostly caused by bacterial plaque, calculus, food impaction, bad restorations and bite trauma. Due to the characteristics of the subgingival micro-ecological environment, a large number of highly virulent periodontal pathogenic bacteria, such as Bacteroides gingivalis, Bacteroides intermedia, and spirochetes, grow in the subgingival plaque, which aggravates and spreads the inflammation of the gums, resulting in Periodontal pocket formation and alveolar bone resorption ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/38A61P1/02A61K35/28
CPCA61K38/38A61K9/0019A61K35/28A61K2300/00
Inventor 葛啸虎陈海佳王一飞陈育广王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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