Method of preparing stem cell line for excreting insulin by using slow virus vectors of coding multiple exogenous gene

A technology of exogenous genes and stem cell lines, applied to cells modified by introducing foreign genetic materials, using vectors to introduce foreign genetic materials, recombinant DNA technology, etc., can solve problems such as inability to achieve stable cell transfection, and improve insulin expression level, high transfection efficiency, and simple construction method

Inactive Publication Date: 2008-01-09
CHINA JAPAN FRIENDSHIP HOSPITAL
View PDF0 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the past, adenovirus was used to simultaneously transfect the three insulin-regulated genes

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method of preparing stem cell line for excreting insulin by using slow virus vectors of coding multiple exogenous gene
  • Method of preparing stem cell line for excreting insulin by using slow virus vectors of coding multiple exogenous gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0027] A method for preparing an insulin-secreting stem cell line using a lentiviral vector encoding multiple foreign genes, comprising the steps of:

[0028] 1) Construction of lentiviral vectors encoding Neomycin, MafA, NeuroD and PDX-1 genes

[0029] First, the lentiviral vector plasmid pWPTS-GFP was double digested with BamH1 and Sal I enzymes, and after releasing GFP, the vector pWPTS fragment was recovered; then the plasmids containing the genes of Neomycin, MafA, NeuroD1 and PDX-1 were passed through BstB1 and After digestion with Bgl II, Xbal I and Not I, EcoR I and Xbal I, and EcoRI restriction endonucleases, use 0.7% agarose gel electrophoresis analysis to recover the target gene fragments; Ligated into the vector pWPTS fragment, constructed into lentiviral vectors encoding Neomycin, MafA, NeuroD and PDX-1 genes respectively;

[0030] 2) Packaging of lentiviral particles

[0031] ①Amplification of packaging plasmid:

[0032] The lentiviral packaging plasmids pMD2G...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

Production of secretory insulin dry cell system by encoded various exogenous gene slow virus carrier is carried out by copying pWPST, cloning various insulin regulating genes to the carrier, transfecting 293T cell, packing, collecting virus grains, concentrating and transfecting human embryo pancreas dry cell. It's simple, efficient and practical, it can obtain one, two or three kinds of exogenous gene cells, which are in different expression and meet different needs.

Description

technical field [0001] The invention relates to a method for preparing insulin-secreting stem cell lines, in particular to a method for preparing insulin-secreting stem cell lines using lentiviral vectors encoding multiple foreign genes. Background technique [0002] Diabetes is one of the major diseases that seriously endanger human health, and has become the third major disease after cardiovascular diseases and tumors. In recent years, the incidence of diabetes has been increasing, and the onset is getting younger. According to literature reports: the diabetic patient of the whole world has surpassed 230 million, and with the speed increase of 2 million people every year. [0003] Diabetes patients in my country have exceeded 40 million and are expected to reach 60 million by 2010. Diabetic patients have disorders of glucose metabolism due to insulin production disorders or insulin failure to exert physiological functions, resulting in microvascular disease, causing a se...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N15/86C12N5/10
Inventor 门秀丽张文健娄晋宁
Owner CHINA JAPAN FRIENDSHIP HOSPITAL
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap