Method for enhancing secretion function of mesenchymal stem cells and application

A mesenchymal stem cell and stem cell technology, applied in the field of enhancing the secretory function of mesenchymal stem cells, can solve the problems of high price, changing the genome of cells, etc., and achieve the effects of enhancing repair function, enhancing secretion function, and promoting stem cell secretion function

Pending Publication Date: 2020-12-08
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the inventors found that these methods are still in the research stage, and some factors are relatively expensive, and the appropriate concentration and treatment time need to be further explored; hypoxic pretreatment requires special culture equipment, and the conditions are relatively high; Genome, its safety needs to be further clarified

Method used

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  • Method for enhancing secretion function of mesenchymal stem cells and application
  • Method for enhancing secretion function of mesenchymal stem cells and application
  • Method for enhancing secretion function of mesenchymal stem cells and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Cytokine chip detection of the effect of HA pretreatment on the secretion function of stem cells

[0045] S1. The same number of UCMSCs were cultured in normal medium or medium supplemented with 0.3mg / ml HA for 5 days, and the medium was changed on the 3rd day, and the culture was continued after the medium change.

[0046] S2. After 5 days, the two groups of cells were replaced with 13 ml of serum-free basal medium, cultured in the incubator for 48 hours, and the supernatants of the two groups were collected.

[0047] S3. After the cell supernatant was centrifuged at 1500×g, filtered through a 0.22 μM filter, centrifuged at 5000×g for 50 min at room temperature through a 3KDa ultrafiltration centrifuge tube, and the concentrated supernatant was collected for subsequent experiments.

[0048] S4. The concentrated cell supernatants of the two groups were sent to Raybio Biotechnology Co., Ltd. for cytokine chip (Raybiotech, GSH-CAA-440) detection, and scatter pl...

Embodiment 2

[0050] Example 2: ELISA method to verify the results of the cell chip

[0051] The process of collecting the concentrated supernatant was as described in Example 1. The supernatants of the control group and the HA treatment group were sent to Beijing North Biotechnology Research Institute Co., Ltd. for ELISA testing, and the verification factors included HGF, VEGF, EGF, and SCF.

[0052] The results show that: if image 3 As shown, after HA treatment, the secretion of cytokines HGF, VEGF, EGF, and SCF increased, which was consistent with the chip results.

Embodiment 3

[0053] Example 3: Ovarian culture in vitro to detect the effect of stem cell supernatant on PI3K-AKT pathway activation after HA pretreatment

[0054] S1. The process of collecting the supernatant of mesenchymal stem cells is as described in Example 1.

[0055] S2. Prepare ovarian dissection solution. Ovarian dissection solution containing 10% FBS and 1% penicillin-streptomycin was prepared with L15 medium.

[0056] S3. Prepare the ovaries for culture. The culture media of the control group, VCD (30uM) group, VCD+MSC-CM group and VCD+HA / MSC-CM group were prepared, and the embedded culture chamber was placed in a 6-well plate culture medium (1.5ml per well) to equilibrate for 2h.

[0057] S4. Under a stereoscope, the ovaries of suckling mice at postnatal day 4 (PD4) were isolated in the freshly prepared dissecting fluid. Gently peel off the surrounding tissue and capsule of the ovary, and be careful not to damage the ovarian tissue.

[0058] S5. Wash the obtained ovaries th...

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Abstract

The invention provides a method for enhancing the secretion function of mesenchymal stem cells and application, and belongs to the technical field of biology. Research shows that by using Hyaluronic acid sodium (HA) to pretreat umbilical cord mesenchymal stem cells in vitro, the secretion function of the umbilical cord mesenchymal stem cells can be significantly promoted, secretion of HGF, SCF, EGF, VEGF and other factors is promoted, activation of a PI 3K-AKT signal channel is further promoted, and the repair effect on damage is enhanced. The method is simple, convenient, low in cost and highin safety, and therefore the method HAs good practical application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method and application for enhancing the secretory function of mesenchymal stem cells. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Stem cells have great development potential and clinical application value in the fields of tissue engineering and disease treatment. Among them, mesenchymal stem cells (MSCs) widely exist in the connective tissue of the whole body and the interstitium of organs. a type of adult stem cell. In recent years, it has become one of the important research directions in the field of biomedicine. [0004] Existing studies have shown that me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K35/28A61P15/08
CPCC12N5/0668A61K35/28A61P15/08C12N2501/905A61P15/00C12N5/0665
Inventor 赵世斗焦文林秦莹莹陈子江米欣燕涛
Owner SHANDONG UNIV
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