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Stem cell freezing medium and stem cell freezing method

A cryopreservation method and stem cell technology, which is applied in the field of stem cell cryopreservation solution, can solve the problems of damage, affecting the treatment effect, and the activity rate of dental pulp stem cells not reaching the ideal level, so as to reduce damage, avoid pollution, and increase the activity rate. Effect

Inactive Publication Date: 2015-11-25
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, the formula of cell cryopreservation solution for dental pulp stem cells is 10% DMSO + 90% FBS. This cell cryopreservation solution will cause certain damage to dental pulp stem cells during the process of preservation or transportation, making dental pulp stem cells The survival rate has not reached the ideal level, which affects its therapeutic effect

Method used

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  • Stem cell freezing medium and stem cell freezing method
  • Stem cell freezing medium and stem cell freezing method

Examples

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Embodiment 1～3

[0034] Example 1-3 Preparation of Dental Pulp Stem Cells

[0035] (1) Isolation and culture of dental pulp stem cells:

[0036] With the consent of the family members, the clinically extracted deciduous teeth of children aged 6-10 were collected due to retention, and no dental pulp disease and necrosis were required. Immediately after pulling it out, put it in a PBS centrifuge tube containing double antibodies (100 μg / mL penicillin, 100 μg / mL streptomycin) when it is cold at 4°C, and take it out for use within 24 hours.

[0037] Take out the tooth, rinse it three times repeatedly with PBS solution containing double antibody, wrap the tooth in sterile gauze and crack the tooth with forceps to expose the pulp tissue; use sterile tweezers to grasp the pulp tissue, and excise the 1mm tooth at the root tip marrow tissue. According to the size of the pulp tissue, cut the pulp tissue into 1mm with ophthalmic curved scissors 3 , placed in a 50mL centrifuge tube. Add 10 times the v...

Embodiment 4

[0052] Example 4 Preparation of Dental Pulp Stem Cell Injection

[0053] (1) Isolation and culture of dental pulp stem cells

[0054] 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 3g / LI type collagenase, seal and mix well, transfer to a constant temperature air shaker, digest for about 10-20min at 37°C and 200R, add an equal volume of complete The medium was digested, 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 1000rpm for 5min. The precipitate was resuspended in serum-free medium, ...

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Abstract

The invention relates to the field of biotechnology, and in particular relates to a stem cell freezing medium and a stem cell freezing method. The stem cell freezing medium consists of DMSO, hydroxyethyl starch and a serum-free medium. The stem cell freezing medium provided by the invention can be used for remarkably reducing injury of the freezing medium on cells, so as to significantly improve the survival rate of the stem cells; and by adopting the serum-free medium, potential pollution caused by serum is avoided.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a stem cell cryopreservation solution and a stem cell cryopreservation method. 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 and mind. [0003] The main purpose...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 陈海佳王一飞葛啸虎冯德龙马岩岩
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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