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Gingival mesenchymal stem cell exosome preparation and application thereof

A technology of stem cells and exosomes, applied in the field of gingival mesenchymal stem cell exosome preparations, can solve the problems of inability to completely recover and easy recurrence of psoriasis.

Pending Publication Date: 2020-05-12
GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the treatment of psoriasis can only alleviate the condition and is easy to relapse, and cannot be completely cured

Method used

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  • Gingival mesenchymal stem cell exosome preparation and application thereof
  • Gingival mesenchymal stem cell exosome preparation and application thereof
  • Gingival mesenchymal stem cell exosome preparation and application thereof

Examples

Experimental program
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Embodiment 1

[0045]Example 1 Isolation, cultivation and detection of gingival mesenchymal stem cells

[0046] (1) Culture and passage of gingival mesenchymal stem cells

[0047] ①Cultivation: Soak the gingival tissue with povidone iodine for 30s in the ultra-clean workbench, wash the gingival tissue 3 times with PBS, and cut the washed gingival tissue to 3mm with a scalpel 2 size. Transfer the minced tissue pieces to an EP tube containing 4 mg / mL type IV collagenase, and digest with shaking at 37°C for 2 hours. After 2h, centrifuge at 1000r / min for 5min, discard the supernatant, resuspend with α-MEM complete medium containing 10% FBS, and transfer to a 6cm culture dish at 37°C, 5%CO 2 cultivated under conditions. Let it stand for the first 3 days, change the medium on the 4th day, and then change the medium every 3 days. Observe the microscopic morphology of primary gingival mesenchymal stem cells on the first day (day1), third day (day3), fifth day (day5) and seventh day (day7) under ...

Embodiment 2

[0073] Example 2 Extraction and detection of exosomes derived from gingival mesenchymal stem cells

[0074] (1) Exosomes derived from gingival mesenchymal stem cells were extracted by ultracentrifugation

[0075] Select the second-generation gingival mesenchymal stem cells in a good growth state. When the cell confluence reaches 90%, use exosome-free serum conditioned medium (purchased from ScienCell Company), and collect the cell culture supernatant after continuous culture for 48 hours, 300g Centrifuge for 10min, take supernatant, and remove cells; centrifuge at 2000g for 10min at 4°C, take supernatant, and remove dead cells; centrifuge for 30min at 10,000g at 4°C, take supernatant, remove cell debris; filter supernatant with 0.22μm filter, Centrifuge at 100,000g at 4°C for 70min, discard the supernatant, keep the precipitate, add 10mL PBS to resuspend; centrifuge at 100,000g at 4°C for 70min, discard the supernatant, keep the precipitate, the precipitate is exosomes derived...

Embodiment 3

[0082] The establishment of embodiment 3 psoriasis mouse model

[0083] Eighteen SPF female C57BL / 6 mice (6-8 weeks old, body weight 16-18 g) were selected. Carefully shaved off the central area of ​​the mouse's back, and then removed the superficial vellus hair with a mild depilatory cream. From the second day after depilation on the back of mice (62.5mg / cm 2 ) and smear 5% imiquimod cream, and the mice were continuously modeled for 6 days.

[0084] The successfully established psoriasis mouse models were randomly divided into 3 groups: negative control group (NC), positive drug treatment group (CP), and gingival mesenchymal stem cell exosome treatment group (GMSCs-exo) group. Group 6. 100 μL of PBS was applied to the lesions in the NC group, and 20 mg / cm2 was applied to the lesions in the CP group. 2 The positive drug (clobetasol propionate ointment), GMSCs-exo group applied 100 μL of GMSC-derived exosomes (1 mg / mL) to the lesion. Once a day, for 5 consecutive days, obs...

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Abstract

The invention discloses a gingival mesenchymal stem cell exosome preparation and application thereof, and belongs to the technical field of biology. The invention discloses an application of an exosome derived from gingival mesenchymal stem cells in preparation of a medicine for treating treatment psoriasis. The gingival mesenchymal stem cell exosome preparation disclosed by the invention can be locally applied to a psoriasis diseased region and can relieve or inhibit inflammatory cell infiltration and inflammatory factor expression of the diseased region, so that the effects of treatment anddisease relief of diseases are achieved; the exosome preparation can also be used as a psoriasis drug additive or a combined drug preparation for psoriasis treatment.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to a gingival mesenchymal stem cell exosome preparation and its application. Background technique [0002] Gingival mesenchymal stem cells (GMSC) are precursor cells isolated from human gingival tissue with stem cell characteristics, which have the functions of self-renewal, multilineage differentiation and immune regulation. Most importantly, gingival mesenchymal stem cells can secrete extracellular vesicles with a diameter of about 30-150 nm, which are often called exosomes. Exosomes are widely found in animal and plant cells, but their functions are closely related to cell types and types. Its core functions mainly include intercellular communication, tissue repair and regeneration, immune regulation, body development, and disease diagnosis. At present, gingival mesenchymal stem cells have been confirmed to have a certain role in autoimmune diseases, neurological di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P17/06C12N5/0775
CPCA61K35/28A61K9/0014A61P17/06C12N5/0668
Inventor 曾今诚叶子瑜李妍芸林碧华梁艳芳杨维青
Owner GUANGDONG MEDICAL UNIV
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