Exosome generated by mesenchymal stem cells treated by IL18, IL12 and IL15 and antiviral application

A technology of stem cells and exosomes, applied in the field of biomedicine, can solve the problems of poor treatment effect and lack of treatment methods for viral pneumonia, and achieve the effects of good uniformity, reduced damage and high particle content.

Pending Publication Date: 2022-05-31
SHENZHEN BEIKE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The technical problem to be solved by the present invention is that the current

Method used

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  • Exosome generated by mesenchymal stem cells treated by IL18, IL12 and IL15 and antiviral application
  • Exosome generated by mesenchymal stem cells treated by IL18, IL12 and IL15 and antiviral application
  • Exosome generated by mesenchymal stem cells treated by IL18, IL12 and IL15 and antiviral application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach 1

[0046] Embodiment 1 Pretreatment of Human Umbilical Cord Mesenchymal Stem Cells with Cytokine Composition

[0047] Isolation and culture of mesenchymal stem cells:

[0048] S1. Under the supervision of the Medical Ethics Committee, fresh umbilical cords were obtained from healthy donors for the separation of Wharton's jelly, the Wharton's jelly was cut into pieces, and the adherent method was used to culture the primary cells in a serum-free culture system. The acquisition and expansion of the serum-free medium is: Ultraculture (Lonza) + commercially available MSCs culture factors.

[0049] S2. The primary cells are amplified and frozen in the P2 generation as seed cells.

[0050] In this specification, the "P2 generation" refers to the generation of primary cells in culture flasks and / or cell factories, after two times of confluence reaching 100%, digestion and replanting into culture flasks and / or cell factories.

[0051] S3. Pretreatment of human umbilical cord mesenchyma...

Embodiment approach 3

[0069] Embodiment 3 In vitro immunosuppressive test of exosomes produced by human umbilical cord mesenchymal stem cells before and after pretreatment

[0070] Co-cultivating mesenchymal stem cells with human peripheral blood mononuclear cells in vitro is an important experimental method for detecting the immunosuppressive ability of mesenchymal stem cells in vitro.

[0071] In the present invention, after human peripheral blood mononuclear cells are labeled with CFSE and activated, they are mixed with exosomes produced by ordinary human umbilical cord mesenchymal stem cells and exosomes produced by pretreated human umbilical cord mesenchymal stem cells (pretreated During the treatment, the combination types used in the cytokine composition include: factor combination ① IL18, IL12, IL15; factor combination ② IFN-γ, IL21, IL23) for co-cultivation, it can be seen that after 80 hours of co-cultivation, after factor combination ① The immunosuppressive effect of exosomes produced by...

Embodiment approach 4

[0075] Embodiment 4 Exosomes produced by human umbilical cord mesenchymal stem cells pretreated with different factor compositions to treat viral pneumonia mouse model

[0076] H1N1 virus-induced acute lung injury in mice is a classic animal model that mimics human viral or autoimmune acute pneumonia. In this model, the acute immune response plays a dominant role in mediating lung injury, and various immune cells (T cells, B cells, macrophages, NK cells) and inflammatory factors (IFN-γ, IL-1β , IL6, TNF-α, etc.) are all involved in the course of acute pneumonia, and the lungs of mice infected with the virus will have a large number of immune cell infiltration, red blood cell precipitation, and lung structural damage.

[0077] In the present invention, the exosomes produced by ordinary human umbilical cord mesenchymal stem cells and the exosomes produced by human umbilical cord mesenchymal stem cells pretreated by factor combination ① have the characteristics of immunosuppressi...

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Abstract

The invention discloses exosomes generated by mesenchymal stem cells treated by IL18, IL12 and IL15 and antiviral application of the exosomes, and relates to the technical field of biological medicines. The exosome is generated by pretreating mesenchymal stem cells with a cytokine composition, and the cytokine composition contains at least two cytokines of IL18, IL12 and IL15. Compared with exosomes secreted by human mesenchymal stem cells which are not pretreated, the exosome of the mesenchymal stem cells with the enhanced antiviral pneumonia activity, provided by the invention, has better antiviral pneumonia activity so as to be used for treating immune inflammatory reaction caused by viral pneumonia and reducing the damage of the inflammatory reaction to lung tissues; the effect of repairing tissues and organs is achieved; the exosome secreted by the pretreated human mesenchymal stem cells has the characteristics of better homogeneity, higher immunoregulatory protein content, higher exosome particle content and the like; the compound can be applied to preparation of drugs for treating viral pneumonia or immune inflammation caused by viral pneumonia.

Description

technical field [0001] The present invention relates to the technical field of biomedicine, in particular to exosomes produced by IL18, IL12, and IL15-treated mesenchymal stem cells and their antiviral applications. The exosomes have enhanced antiviral pneumonia activity. Background technique [0002] Since the end of 2019, the viral pneumonia caused by the novel coronavirus that has ravaged the world has attracted the attention of the medical community around the world. This type of virus mutates rapidly. Once infected with the disease, the disease progresses faster. Patients with underlying diseases are infected After the new coronavirus, it is easy to develop severe viral pneumonia and cause acute respiratory distress syndrome, cytokine storm in the body, etc. At present, the treatment methods for viral pneumonia are generally based on supportive treatment, antiviral treatment, immunotherapy, and glucocorticoid treatment, while respiratory support and prevention of compli...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28A61P11/00A61P31/12
CPCC12N5/0668A61K35/28A61P11/00A61P31/12C12N2501/2312C12N2501/2315C12N2501/2318
Inventor 廖延傅泽钦杨玉林伍世铎刘沐芸胡隽源蔡车国
Owner SHENZHEN BEIKE BIOTECH
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