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Treatment of type 2 diabetes mellitus with gene modified MSCs

A gene and gene-encoding technology, applied in the preparation method and application, as well as in the field of drugs for treating type 2 diabetes and recombinant stem cells, can solve problems such as inability to treat diabetes, and achieve the effects of improving sensitivity, long-term action, and reducing blood sugar and blood lipid levels Effect

Active Publication Date: 2019-12-31
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some oral antidiabetic drugs mainly achieve the purpose of lowering blood sugar by stimulating the secretion of insulin by the remaining islet cells, and insulin can only be used as a supplementary or replacement therapy for relative or absolute lack of insulin, neither of which can treat diabetes from the source by targeting the pathogenesis

Method used

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  • Treatment of type 2 diabetes mellitus with gene modified MSCs
  • Treatment of type 2 diabetes mellitus with gene modified MSCs
  • Treatment of type 2 diabetes mellitus with gene modified MSCs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1 provides a method for preparing recombinant mesenchymal stem cells, comprising:

[0048] 1. Isolation and identification of adipose-derived mesenchymal stem cells

[0049] Human adipose-derived mesenchymal stem cells and adipose tissue were provided by the Plastic Surgery Department of Peking Union Medical College Hospital. The samples were obtained and volunteers signed an informed consent. The fat donors are healthy people aged 25-35 without HIV, HBV and HCV infection. The fat sample is the discarded fat during liposuction, which is sucked out and stored in a sterile, airtight disposable drainage bag, stored at low temperature and transported to the laboratory for the isolation of primary adipose-derived mesenchymal stem cells. Prepare collagenase type II 0.2%, a large amount of HBSS+2% P / S, wash the fat with HBSS equal to the volume of fat for 5 times, and wash away the blood components. Add collagenase solution equal to the volume of fat in each tube,...

Embodiment 2

[0064] Example 2 Determination of biochemical indicators

[0065] The obese diabetic mouse model was established by feeding the mice with high fat for 16 weeks, and the blood glucose concentration in the tail vein blood was detected with a blood glucose meter. Blood samples were collected by heart blood collection, put into EDTA anticoagulant tubes, and centrifuged at 2000 rpm at 4°C for 15 min. The supernatant was taken for the detection of blood indicators. 4 μl of plasma was taken to detect the insulin content with a kit (Mercodia, 10-1247-01). 10 μl of plasma was taken to detect triglyceride content with a kit (Sigma, TR0100). Take 20 microliters and send it to the hospital for liver function testing.

[0066] By feeding mice with high fat for 16 weeks, an obese diabetic mouse model was established, and the weight of the mice increased significantly, reaching obesity, such as image 3 as shown in a. Detect that the blood glucose level of mice fed with high-fat feed is...

Embodiment 3

[0070] Example 3 Detection of MSCs survival and distribution in vivo

[0071] The survival and distribution of MSCs in vivo are very important for their therapeutic effects. Therefore, using Akaluc luciferase to react with luciferin to generate bioluminescent signals can detect the position, distribution and number of cells in the body. Taking MSCs cells as an example, by constructing a lentivirus containing the gene encoding Akaluc luciferase, and infecting MSCs cells with the lentivirus, MSC-Akaluc cells (that is, MSC cells capable of expressing Akaluc luciferase) can be obtained. Express luciferase, which produces bioluminescence when Akaluc substrate is added, by The strength of the signal captured by Lumina II reflects the amount of cells.

[0072] Therefore, we injected MSC-Akaluc cells into mice fed with conventional diet and high-fat diet, and injected Akaluc substrate intraperitoneally, and observed the bioluminescent signal 6 hours after injection, and found that ...

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Abstract

The invention relates to the technical field of biology and relates to treatment of type 2 diabetes mellitus with gene modified MSCs. The invention provides a recombinant stem cell which can overexpress an Exendin-4 protein. Medicines for treating obesity, fatty liver and type 2 diabetes mellitus can be prepared by utilizing the recombinant stem cell. The recombinant stem cell can be used for remarkably reducing the level of blood glucose and blood lipids and promoting the sensitivity of insulin. The treatment effect can be maintained for one month or more after a course of treatment, and therecombinant stem call can be used for treating metabolic diseases including obesity, fatty liver and type 2 diabetes mellitus, is safe and effective and has a long acting time.

Description

technical field [0001] The invention relates to the field of biotechnology, relates to gene-modified MSCs for treating type 2 diabetes, and in particular to a recombinant stem cell, a preparation method and use thereof, and a medicine for treating type 2 diabetes. Background technique [0002] Diabetes is one of the most serious and critical health problems facing the world today. In 2017, about 425 million adults worldwide suffered from diabetes, and more than 4 million people died of diabetes and its complications every year, and the total medical expenditure of diabetes reached 727 billion US dollars. Despite continued efforts, the number of people with diabetes continues to grow and is expected to reach 629 million by 2045. The current anti-diabetic drugs can only relieve symptoms and delay the progression of the disease, but cannot fundamentally cure it, seriously affecting the quality of life of patients. Therefore, it is urgent to prevent and completely cure diabete...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/867A61K35/28A61P3/10
CPCA61K35/28A61P3/10C07K14/57563C12N15/86C12N2740/15043
Inventor 杜亚楠张元元高爽
Owner TSINGHUA UNIV
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