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Production method and application of an exosome secretion inducer, induction medium, and exosomes using the same

A technology of exosomes and inducers, applied in the application field of medicine, can solve the problems of limited production, achieve the effect of promoting secretion, increasing production, and simple ingredients

Active Publication Date: 2022-06-03
GUIDON PHARM INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing exosome acquisition method is mainly cell autocrine, but its output is very limited

Method used

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  • Production method and application of an exosome secretion inducer, induction medium, and exosomes using the same
  • Production method and application of an exosome secretion inducer, induction medium, and exosomes using the same
  • Production method and application of an exosome secretion inducer, induction medium, and exosomes using the same

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Experimental program
Comparison scheme
Effect test

Embodiment 1

The present embodiment is to adapt to high-yield exosome production, a kind of exosome secretion for efficient production of exosome is provided

induction medium.

[0038] The exosome secretion-inducing cultures formulated for the high-efficiency production of exosomes from stem cells (ESCs or iPSCs, etc.) of the present invention.

A nutrient medium (named GDEV medium) consisting of: DMEM / F12 (Gibco 11330032) as basal medium and

Exosome secretion inducers. The exosome secretion inducer is a component of the following concentrations: L-64mg / L

Ascorbic acid 2phosphate magnesium salt (L‑ascorbic acid magnesium diphosphate salt, Sigma‑Aldrich

A8960), sodium selenium (sodium selenium, Sigma-Aldrich S5261) at a concentration of 14 μg / L, at a concentration of 543 mg / L

NaHCO

3

(Sigma-Aldrich S5761), Insulin at a concentration of 19.4 mg / L (Insulin, Proceeding I10022).

As contrast, prepare its E8 substratum according to the method disclosed in document 1, be specially: DM...

Embodiment 2

[0065]

[0067]

Embodiment 3

5%CO

2

After 24 hours of incubation in the incubator, the cells were stained with Hoechst 33342 / PI, and the neuronal activity was analyzed by high-content photography.

analysis.

Result is as shown in Figure 6, it is the dead cell ratio histogram of statistics, and wherein NO OGD represents not carrying out OGD model,

DPQ means adding DPQ (PARP-1 (inhibitor of poly(ADP-ribose) polymerase-1) to cells treated with DPQ confirmed

Under the influence of ischemia, apoptosis can be reduced. It can be seen that relative to the control group (OGD group), experimental group 2 (ie MSC group) and

The proportion of dead cells in experimental group 3 (ie, E8 group) was significantly reduced. At the same concentration of exosomes, experimental group 1 (ie, iPSC)

The proportion of dead cells in EV group) was significantly lower than that in experimental group 3 (ie, E8 group) and experimental group 2 (ie, MSC group), indicating that the GDEV medium

The exosomes obtained by inducin...

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Abstract

The invention discloses an exosome secretion inducer, an induction medium, and a production method and application of an exosome using the same, belonging to the field of biotechnology. The exosome secretion induction medium provided by the present invention can only contain basal medium and four kinds of supplements: L-ascorbic acid or its salt, selenium or its salt, NaHCO 3 And Insulin, its composition is simple, but can significantly increase the production of exosomes secreted by stem cells, which can significantly reduce the production cost of exosomes; the production of exosomes can more effectively protect neurons from damage and improve aging cells Happening.

Description

An exosome secretion inducer, induction medium and production of exosomes using the same Production method and application technical field The invention belongs to biotechnology field, be specifically related to a kind of exosome secretion inducer and induction medium and application The production method and application of exosomes thereof, especially relate to the preparation of exosomes to improve the condition of senescent cells and / or protect nerves. Metapharmaceutical applications. Background technique [0002] Exosomes are specifically secreted small membrane vesicles (30-150 nm) containing complex RNAs and proteins, which are involved in cellular Intercellular communication, regulating the normal function and damage repair of various tissues and organs. Since exosomes derived from different tissues have their specific protein molecules and key components for their functioning Therefore, exosomes derived from stem cells have important therapeutic effects of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/074C12N5/0735A61K35/28A61P39/06
CPCC12N5/0606C12N5/0696A61K35/28A61P39/06C12N2500/38C12N2500/25C12N2500/05
Inventor 张建民王靓曲典林埕宇
Owner GUIDON PHARM INC