Dental pulp mesenchymal stem cell cryopreservation solution and cryopreservation method thereof

A technique of stem cells and cryopreservation method is applied in the field of cryopreservation of dental pulp mesenchymal stem cells, which can solve the problems of affecting the clinical utilization of stem cells, unable to guarantee the vitality of stem cells, and damage of dental pulp stem cells, so as to maintain biological characteristics, The effect of reducing damage and improving cell viability

Inactive Publication Date: 2017-02-22
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, these cryopreservation solutions will inevitably cause certain damage to dental pulp stem cells during the cryopreservation process of cells, and the vitality of stem cells cannot be guaranteed, and the cell proliferation rate is low, which affects the clinical utilization of stem cells

Method used

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  • Dental pulp mesenchymal stem cell cryopreservation solution and cryopreservation method thereof
  • Dental pulp mesenchymal stem cell cryopreservation solution and cryopreservation method thereof
  • Dental pulp mesenchymal stem cell cryopreservation solution and cryopreservation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] (1) Culture of dental pulp mesenchymal stem cells and collection of conditioned medium

[0041] (1) Culture of dental pulp mesenchymal stem cells

[0042] The healthy teeth that fell off and were extracted from people under 30 years old were collected, the dirt on the tooth surface was cleaned with PBS, the teeth were cracked with forceps, and the pulp tissue was exposed; the pulp tissue was grasped with sterile forceps. Cut into pieces, place in a 50mL centrifuge tube, add 10 times the volume of 3-5g / L type I collagenase, seal and mix well, transfer to a constant temperature air shaker, digest at 37°C and 200R for about 10-20min, add an equal volume The complete medium in the complete medium was used to terminate the digestion, and the discrete cell aggregates were pipetted repeatedly, filtered through a 70 μm cell mesh to obtain single discrete cells, and centrifuged at 1000 rpm for 5 min. Discard the supernatant, wash the pellet once with 30mL PBS, and centrifuge at...

Embodiment 2

[0059] (1) Culture of dental pulp mesenchymal stem cells and collection of conditioned medium

[0060] (1) Culture of dental pulp mesenchymal stem cells

[0061] The healthy teeth that fell off and were extracted from people under 30 years old were collected, the dirt on the tooth surface was cleaned with PBS, the teeth were cracked with forceps, and the pulp tissue was exposed; the pulp tissue was grasped with sterile forceps. Cut into pieces, place in a 50mL centrifuge tube, add 10 times the volume of 3-5g / L type I collagenase, seal and mix well, transfer to a constant temperature air shaker, digest at 37°C and 200R for about 10-20min, add an equal volume The complete medium in the complete medium was used to terminate the digestion, and the discrete cell aggregates were pipetted repeatedly, filtered through a 70 μm cell mesh to obtain single discrete cells, and centrifuged at 1000 rpm for 5 min. Discard the supernatant, wash the pellet once with 30mL PBS, and centrifuge at...

Embodiment 3

[0078] (1) Culture of dental pulp mesenchymal stem cells and collection of conditioned medium

[0079] (1) Culture of dental pulp mesenchymal stem cells

[0080] The healthy teeth that fell off and were extracted from people under 30 years old were collected, the dirt on the tooth surface was cleaned with PBS, the teeth were cracked with forceps, and the pulp tissue was exposed; the pulp tissue was grasped with sterile forceps. Cut into pieces, place in a 50mL centrifuge tube, add 10 times the volume of 3-5g / L type I collagenase, seal and mix well, transfer to a constant temperature air shaker, digest at 37°C and 200R for about 10-20min, add an equal volume The complete medium in the complete medium was used to terminate the digestion, and the discrete cell aggregates were pipetted repeatedly, filtered through a 70 μm cell mesh to obtain single discrete cells, and centrifuged at 1000 rpm for 5 min. Discard the supernatant, wash the pellet once with 30mL PBS, and centrifuge at...

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Abstract

The invention relates to the technical field of cell culture, and particularly relates to a dental pulp mesenchymal stem cell cryopreservation solution and a cryopreservation method thereof. The cryopreservation solution is a conditioned medium containing DMSO, FBS and dental pulp mesenchymal stem cells; a preparation method of the dental pulp mesenchymal stem cell conditioned medium includes the steps of conducting cell culture of dental pulp mesenchymal stem cells by using a cell culture medium, and after cell confluence degree reaches 80%-90%, collecting supernatant in the cell culture medium to obtain the dental pulp mesenchymal stem cell conditioned medium. The cryopreservation solution is adopted for cryopreservation of the dental pulp mesenchymal stem cells, the damage effect on the dental pulp mesenchymal stem cells by the cryopreservation solution is thus greatly reduced, cell viability of the dental pulp mesenchymal stem cells in cryopreservation process and thawing and waking process are improved, and biological characteristics of the dental pulp mesenchymal stem cells can be better maintained.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a cryopreservation solution of dental pulp mesenchymal stem cells and a cryopreservation method thereof. Background technique [0002] Periodontitis is a chronic infectious disease involving four types of periodontal supporting tissues (gingiva, periodontal ligament, alveolar bone, and cementum), often leading to inflammatory destruction of periodontal supporting tissues. It is more common after the age of 35. Common causes are bacterial plaque, calculus, traumatic occlusion, food impaction, bad restorations, and mouth breathing. Periodontitis has four major characteristics, namely the formation of periodontal pockets, inflammation of the pocket wall, alveolar bone resorption, and gradual loosening of teeth. Severe periodontitis can cause tooth loss, leading to poor masticatory function and indigestion and gastrointestinal diseases, affecting the health of the whole body a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02C12N5/0775
CPCA01N1/0221C12N5/0664
Inventor 葛啸虎陈海佳王一飞冯德龙张维敏
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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