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Applications of exosome from mesenchymal stem cells

A technology of mesenchymal stem cells and exosomes, applied in medical preparations containing active ingredients, sensory diseases, unknown raw materials, etc., can solve the problems of clinical application of tumorigenic risks, low survival rate of transplantation, difficulties in storage and transportation, etc., to achieve Improve retinal function, reduce inflammation, and reduce retinal damage

Active Publication Date: 2016-01-27
天津医科大学眼科医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the difficulties in storage and transportation of MSC cell products, the low survival rate of transplantation and the possible risk of tumorigenicity in long-term clinical application, its clinical application is limited. Therefore, it is necessary to find a simple, safe and effective MSC-based cell-free therapy is very important

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  • Applications of exosome from mesenchymal stem cells
  • Applications of exosome from mesenchymal stem cells
  • Applications of exosome from mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Isolation, purification and identification of exosomes derived from human umbilical cord mesenchymal stem cells (hucMSC)

[0045] 1. A method for separating and purifying exosomes derived from human umbilical cord mesenchymal stem cells, comprising the steps of:

[0046] (1) Isolation and culture of human umbilical cord mesenchymal stem cells: fresh umbilical cords of sterile newborns were taken, washed repeatedly with phosphate buffer saline (PBS), and cut into tissue pieces with a diameter of about 1-2 mm; treated with type 2 collagenase and After trypsin digestion, centrifuge the supernatant, take the cell pellet and put it into a culture bottle, use DMEM / F12 medium containing 10% fetal bovine serum, 5% CO 2 , Culture in saturated humidity at 37°C; remove non-adherent cells, and subculture after 80% fusion of adherent cells with 0.25% trypsin;

[0047] (2) Collection of conditioned medium for mesenchymal stem cells: take 3-5 generations of mesenchymal stem cells and...

Embodiment 2

[0052] Example 2: Observation of curative effect of hucMSC-exosome on mouse retinal injury model

[0053] (1) Model establishment: 6-8 weeks old healthy female C57BL / 6 mice of SPF grade (body weight 15-18 g) were included in the experiment after examination confirmed that the refractive interstitium was clear and the fundus was normal. The husbandry practices followed the Society for Research in Vision and Ophthalmology Standards for the Care and Use of Animals in Ophthalmology and Vision Research. The mice were randomly divided into normal group, treatment group and injury control group. The mice in the normal group did not receive any treatment. The mice in the treatment group and the injury control group were anesthetized by intraperitoneal injection of 5% chloral hydrate at a dose of 500 mg / kg, and after the right eye was dilated with compound tropicamide, laser light was performed using a multi-wavelength krypton ion laser machine NOVUSOMNI (Coherent, USA). Condensation...

Embodiment 3

[0061] Example 3: Observation of curative effect of hucMSC-exosome on experimental autoimmune uveitis model in rats

[0062] (1) Model establishment: 6-8 week-old female Lewis rats, 30 μg of interphotoreceptor retinoid-binding protein (IRBP) R16 polypeptide fragment dissolved in phosphate buffered saline (PBS) and an equal volume containing 2.5 mg / ml Mycobacterium tuberculosis The complete Freund's adjuvant (CFA) of H37Ra was mixed in equal volume, and the three-way valve was fully emulsified. 0.2ml was taken from each rat and subcutaneously injected into one hind paw to establish the EAU model.

[0063] (2) Treatment method and grouping: Lewis rats were randomly divided into exosome treatment group and model control group with 6 rats after EAU modeling. The exosome treatment group was injected with 50 μl of PBS containing 30 μghucMSC-exosome daily in the juxtabulbar sub-tenon capsule from the initial stage of the disease for 7 consecutive days, and the corresponding model co...

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Abstract

The invention relates to applications of exosome from mesenchymal stem cells. The concrete applications are applications of exosome from mesenchymal stem cells in preparation of medicines treating ophthalmic diseases. The ophthalmic diseases comprise uveitis, retinal detachment, retinal degeneration, retinal damage caused by various reasons and inflammation. Experiments prove that exosome has nerve nourishing and immunosuppression functions, can relief retina damage caused by various reasons and can reduce inflammatory reactions, therefore retina functions are improved, and a new approach is provided for clinic treatment of ophthalmic diseases.

Description

technical field [0001] The invention belongs to the fields of drug research and development, cell biology and molecular biology. The invention relates to the application of active ingredient exosome in mesenchymal stem cells (mesenchymalstemcell, MSC) in the preparation of drugs for treating ophthalmic diseases. Background technique [0002] It is known that mesenchymal stem cells (MSCs) play a role in the treatment of various tissue injuries and immune diseases. We also demonstrated that MSCs have therapeutic effects in various ophthalmic disease models including retinal ischemia-reperfusion, diabetic retinopathy, uveoretinitis, retinal degeneration, and retinal injury. MSCs can be isolated from several adult tissues (including bone marrow, adipose tissue, cord blood) and expanded ex vivo. However, due to the difficulties in storage and transportation of MSC cell products, the low survival rate of transplantation and the possible risk of tumorigenicity in long-term clinic...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61P27/02
Inventor 张晓敏李筱荣苏畅
Owner 天津医科大学眼科医院
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