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Human umbilical cord-derived mesenchymal stem cell strain stably expressing exogenous GLP-1 gene

A GLP-1, stem cell technology, applied in the direction of cells modified by introducing foreign genetic material, genetic engineering, plant genetic improvement, etc., can solve the problem of long-term continuous effect, short plasma half-life, and limited clinical application of GLP-1 And other issues

Inactive Publication Date: 2015-07-29
SHANGHAI OE BIOTECH CO LTD
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Problems solved by technology

However, because GLP-1 is easily degraded by dipeptidyl peptide IV (DPP-IV) in the human body; and its plasma half-life is less than 2 minutes, continuous intravenous infusion or continuous subcutaneous injection is required to produce curative effect, which greatly limits GLP-1. 1 clinical application
Solve the problem that GLP-1 itself has a short plasma half-life (less than 2 minutes), and it is difficult to continue to function for a long time

Method used

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  • Human umbilical cord-derived mesenchymal stem cell strain stably expressing exogenous GLP-1 gene

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Embodiment 1

[0022] Embodiment one, see figure 1 Shown:

[0023] A method for preparing a human umbilical cord-derived mesenchymal stem cell line stably expressing an exogenous GLP-1 gene, comprising the steps of:

[0024] (1) Isolation and culture of human umbilical cord-derived mesenchymal stem cells: human umbilical cord tissue from normal delivery was collected and separated aseptically, and the umbilical cord tissue was cut into 1cm 3 Add the a-MEM culture solution containing FGFb at a final concentration of 20ng / ml to the tissue block, at 37°C, 5% CO 2 Cultivate in an incubator. After 3 days, human umbilical cord-derived mesenchymal stem cells crawl out from the umbilical cord tissue. When the cells reach 80% fusion rate, collect the cells and set aside;

[0025] (2) Construction of pCDNA3.1-GLP-1 eukaryotic expression vector: select the coding gene part in the sequence according to the GLP-1 sequence information provided on GeneBank, use the Overlap-PCR method to obtain the full l...

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Abstract

The present invention relates to the technical field of gene preparation and cell culture, particularly to a human umbilical cord-derived mesenchymal stem cell strain stably expressing exogenous GLP-1 gene, wherein human umbilical cord-derived mesenchymal stem cells are separated and cloned from human umbilical cord, an eukaryotic expression plasmid pCDNA3.1 is adopted as a vector to transform exogenous GLP-1 gene into the human umbilical cord-derived mesenchymal stem cells, and G418 pressurizing screening culture is performed to obtain the cell strain. With the human umbilical cord-derived mesenchymal stem cell strain stably expressing exogenous GLP-1 gene, the disadvantage that continuous administration of the GLP-1 protein can perform the 2 diabetes mellitus treatment due to easy degradation and short half-life is solved when use of the GLP-1 protein to treat 2 diabetes mellitus.

Description

Technical field: [0001] The invention relates to the technical field of gene preparation and cell culture, in particular to a human umbilical cord-derived mesenchymal stem cell strain stably expressing exogenous GLP-1 gene and a preparation method thereof. Background technique: [0002] Diabetes mellitus (DM) is a metabolic disease characterized by hyperglycemia caused by insufficient insulin secretion and / or impaired insulin action. It is currently the third serious threat to human health after cardiovascular and cerebrovascular diseases and cancer. disease. Diabetes can be divided into insulin-dependent (IDDM, type I), non-insulin-dependent (NIDDM, type II) and gestational diabetes, among which type II patients account for more than 80% of diabetes cases. At present, there are more than 150 million patients with type II diabetes (2Diabetes Mellitus, 2DM) worldwide, and it is estimated that the number of patients with type II diabetes will reach 300 million by 2025. Type ...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/16C12N15/85C12R1/91
Inventor 王树伟
Owner SHANGHAI OE BIOTECH CO LTD
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