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Mesenchymal stem cells for intervention in stress-induced cognitive impairment

A technology of mesenchymal stem cells and cognitive impairment, applied in the application field of mesenchymal stem cells in the treatment of stress cognitive impairment drugs, can solve the problems of unclear mechanism, scarcity of drugs, limited dosage, etc., to improve the characteristics of brain aging , Improve pathological features, reduce the effect of administration times

Active Publication Date: 2022-07-26
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the mechanism of stress cognitive impairment is not clear, and due to the blood-brain barrier in the brain, the dose of drugs that can reach the brain area through conventional administration routes is limited, making the current clinical intervention drugs for stress cognitive impairment extremely scarce

Method used

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  • Mesenchymal stem cells for intervention in stress-induced cognitive impairment
  • Mesenchymal stem cells for intervention in stress-induced cognitive impairment
  • Mesenchymal stem cells for intervention in stress-induced cognitive impairment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Construction of a dual-promoter expression vector with reporter gene (pCDH-CMV-FGF21mut-GLP1Fc)

[0036] The existing vector pCDH-EF1-FGF21-T2A-GLP1 was mutated to the FGF21 gene, the promoter was changed, and a reporter gene was inserted to track the colonization of MSC cells at a later stage. The FGF21 gene was mutated to increase the activity of this factor. The constructed vector was named pCDH-CMV-FGF21mut-GLP1Fc. The constructed vector was uplifted, packaged with lentivirus, and then infected with MSC cells. The experimental steps include:

[0037] (1) Vector: The vector pCDH-CMV-MCS-EF1-copGFP-T2A-Puro (purchased from SBI Company) was double digested with BamHI and NotI, 3ug, 37°C for 3h, and directly purified and recovered.

[0038] (2) Design primers:

[0039] CMV-BamHI-FGF21-F: gctagcgaattcgaatttaaatcggatccGCCACCATGGACTCG

[0040] IgG4Fc-NotI-R: gcgatcgcagatccttcgcggccgcTCATTTACCCGGAGACAGGGAGAGG

[0041] (3) PCR FGF21mut-GLP1Fc fragment, templat...

Embodiment 2

[0051] Example 2 Preparation of recombinant lentivirus carrying FGF21mut-GLP1Fc

[0052] (1) Take out a cryopreserved 293T cell (purchased from ATCC) from liquid nitrogen and quickly put it in a 37°C water bath until the ice cubes disappear, add dropwise to a 15ml centrifuge tube containing 5ml of pre-warmed medium, and centrifuge at 1200rpm for 3min , discard the supernatant, resuspend the cells in 293T medium (10% FBS + 1 mM sodium pyruvate + 2 mM glutamine + 1% non-essential amino acids + DMEM) and inoculate the cells into a 150 mm culture dish at 37°C, 5% CO 2 Saturated humidity culture. During the culture, when the cell confluence reaches more than 90%, carry out subculture, discard the old medium, add 5ml sterile PBS solution, shake gently, wash the cells, discard the PBS solution, and add 2ml 0.25% Trypsin- EDTA digestion solution, digested for 1-2 minutes until the cells were completely digested; added serum-containing medium to terminate the digestion, the cell suspe...

Embodiment 3

[0057] Example 3 Preparation of FGF21mut-GLP1Fc-modified mesenchymal stem cells

[0058] The adipose-derived mesenchymal stem cells were isolated and cultured by the mixed collagenase digestion method. The specific method is as follows: the healthy adult adipose tissue extracted by liposuction was transferred to a 50 mL centrifuge tube, washed with PBS, and centrifuged at 1500 rpm for 5 minutes to obtain the upper layer of fat. organize. Mix I, II and IV collagenases in a ratio of 1:1:1 to prepare a 0.2% mixed collagenase, and add adipose tissue to the mixed collagenase digestion solution at a ratio of adipose tissue:collagenase=1:1, Adipose tissue was digested for 30 minutes in a shaker at 37°C. The digested adipose tissue was immediately added to α-MEM cell culture medium (purchased from Gibco) with 10% FBS, 1500 rpm, centrifuged for 10 minutes, and the cells and tissue pellets were pelleted. Cells were resuspended in α-MEM complete medium, and undigested tissue was remove...

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Abstract

The present invention provides the application of mesenchymal stem cells expressing FGF21 and GLP1 in the preparation of drugs for treating cognitive disorders, especially cognitive disorders caused by stress.

Description

technical field [0001] The present application relates to the field of neurological disease treatment and cell therapy, and specifically, the present application provides the application of a mesenchymal stem cell expressing FGF21 and GLP1 in a drug for the treatment of stress-induced cognitive impairment. Background technique [0002] Stress is a state of physical and mental tension and its response caused by the imbalance between actual or cognitive requirements and the ability to adapt and cope in the process of adapting to a variety of adverse factors in the living environment. The stress response has a wide range of effects on the various systems of the body. Moderate stress can improve people's cognitive ability, enhance the flexibility of thinking, and help to start the body's adaptive response to stressors. Excitation can lead to impairment of the body's cognitive function. [0003] Stress-induced cognitive impairment is first manifested in the impact on information...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/28A61K48/00A61P25/28C12N5/10C07K14/50C07K14/605C07K19/00
CPCA61K35/28A61K48/005A61K48/0008A61K9/0019A61P25/28C12N5/0667C07K14/50C07K14/605C07K2319/30C07K2319/31C07K2319/00C12N2510/00
Inventor 钱令嘉薛冰华段海峰王雪谢方赵云王世达
Owner ACADEMY OF MILITARY MEDICAL SCI
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