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Dental pulp stem cell exosome preparation as well as preparation method and application thereof

A technology of dental pulp stem cells and exosomes, which is applied in the field of preparation and preparation of dental pulp stem cells and exosomes, can solve the problems of difficulty in obtaining BMSCs, and achieves reduction of the risk of teratogenic, carcinogenic and immune rejection by injection. Save and apply handy effects

Inactive Publication Date: 2020-07-31
XI AN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the extraction of bone marrow mesenchymal stem cells is also restricted by ethics to a certain extent, so that it is difficult to obtain a sufficient amount of BMSCs for the repair of TBI in clinical practice.

Method used

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  • Dental pulp stem cell exosome preparation as well as preparation method and application thereof
  • Dental pulp stem cell exosome preparation as well as preparation method and application thereof
  • Dental pulp stem cell exosome preparation as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] This example provides a method for preparing exosomes from dental pulp stem cells, which includes the following steps. In this example, adult dental pulp stem cells (dental pulp stem cells, DPSCs) are selected:

[0048] (1) Extraction of dental pulp stem cells, the extraction method includes the following steps in sequence: extraction of wisdom teeth from the Oral and Maxillofacial Surgery Clinic of Xi'an Jiaotong University, complete, no caries, no pulp disease, periapical disease and periodontal disease . The age of the patients is 13-30 years old, the gender is not limited, and there is no infectious disease such as hepatitis and tuberculosis. Before tooth extraction, disinfect the teeth with Aner Iodine III. After complete removal, disinfect again.

[0049] (2) Isolation of dental pulp stem cells.

[0050] Operate in a clean bench, place the extracted wisdom teeth (use an osteotome to separate the root and crown from the enamel cementum junction) in a sterile 35m...

Embodiment 2

[0059] In this example, the dental pulp stem cell exosomes prepared in Example 1 were identified.

[0060] (1) Conduct transmission electron microscope observation.

[0061] The exosomes of dental pulp stem cells were fixed with 4% PFA (paraformaldehyde solution, purchased from Beijing Suo Laibao Technology Co., Ltd.), and the fixative solution was added to the sample loading net and absorbed for 20 min. Add 100 μl PBS to the parafilm to wash, 1% glutaraldehyde for 5 minutes, ultrapure water for 2 minutes (washing 8 times), put the copper grid on 50 μl uranyl oxalate (pH 7) droplet for 5 minutes, put the copper grid on Put on 50μl methylcellulose-UA droplet for 10min, operate on ice, remove the copper mesh with a stainless steel ring, gently absorb the excess liquid on the filter paper, leaving a thin layer of methylcellulose film, the copper mesh is still stainless steel ring and air dry for 5 min. After drying, the appearance of blue-gold interference fringes indicates tha...

Embodiment 3

[0069] In this example, a traumatic brain injury model was constructed and treated with exosomes from dental pulp stem cells.

[0070] Male adult SD rats with a body weight of 200 g were selected and anesthetized by intraperitoneal injection of 10% chloral hydrate at a dose of 350 mg / kg. Find the target anterior bregma, fix the brain of the rat with a stereotaxic instrument, and prepare the skin for disinfection. In the skin preparation area, the skin was cut along the midline sagittal direction, and the subcutaneous tissue was bluntly dissected to expose the bregma. A skull fenestration hole with a diameter of 2 mm was opened at 1 mm to the left side of the bregma (the motor function localization area of ​​the right hind limb of the rat). At a height of 20 cm, a weight of 20 g was used to vertically hit the brain tissue in the window area.

[0071] Dental pulp stem cell exosome therapy. Immediately after the injury, a microsyringe was used to perform local injection treatm...

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Abstract

The invention discloses a dental pulp stem cell exosome preparation as well as a preparation method and application thereof, and relates to the technical field of biology. Dental pulp stem cells can express neuronal cell surface markers, can be differentiated to nerve cells and promote the proliferation of the nerve cells, so that the dental pulp stem cells can be used for preparing cell preparations for repairing nerve injuries such as traumatic brain injuries (TBI). The dental pulp stem cells are used for replacing traditional embryonic stem cells, neural stem cells or bone marrow stem cellsto be used for treating the nerve injuries, and have the advantages of being rich in source, simple and convenient to obtain, low in immunogenicity and capable of avoiding ethical problems. Dental pulp stem cell exosomes can accurately target recipient cells and are used for preparing nerve repair preparations for the injuries such as the TBI, and then treatment of the nerve injuries such as theTBI by the exosomes is achieved. The dental pulp stem cell exosomes can also be used for preparing neuroinflammation inhibitors.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a dental pulp stem cell exosome preparation, preparation method and application thereof. Background technique [0002] Traumatic brain injury (TBI) refers to changes in local anatomy and pathological structure caused by violent factors acting on the head, leading to hemiplegia, aphasia, mental retardation, even coma, and death in patients, and its morbidity and disability rates are high. With the rapid development of my country's economy and society, industries such as transportation, construction, geological exploration, and highly competitive sports have risen rapidly. At the same time, the incidence of TBI caused by production safety accidents and other accidental injuries has shown a continuous upward trend, reaching 100 / 100,000, and the harm is very serious. After the occurrence of TBI, neuronal cell damage is the direct cause of neurological dysfunction. Since neurons are diff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P25/00A61P25/14A61P29/00C12N5/0775
CPCA61K35/28A61P25/00A61P25/14A61P29/00C12N5/0664C12N2509/00
Inventor 李昂李晔裴丹丹王欣欣
Owner XI AN JIAOTONG UNIV
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