Applications of adiponectin modified islet cells in improvement or enhancement of islet transplantation
A technique for islet cell and islet transplantation, which is applied in the field of biomedicine, can solve the problems such as the number of islets and the lack of function that limit the wide application, and achieves the effect of preserving islet function and improving infection efficiency.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0027] The preparation of embodiment 1 adenovirus
[0028] Adenovirus-mediated adiponectin overexpression system and its controls (Ad-APN-GFP and Ad-GFP) were entrusted to Shanghai Jikai Gene for preparation. Plasmid information such as figure 1 shown. Purified high-concentration Ad-APN-GFP was obtained.
Embodiment 2
[0029] Example 2 Adenovirus-mediated APN Gene Transfer of Islet Cells
[0030] After the traditional method treats the islet cluster with adenovirus, the adenovirus can only infect the outermost 1-2 layers of cells of the islet cluster, while the cells in the middle of the islet cluster are difficult to be infected, and the transfection efficiency is low. In order to improve the transfection efficiency of adenovirus to islets, we first treated the islets with 0.25% Trypsin-EDTA solution to disperse them into single cells. Then, we treated dispersed islet cells (60000 cells / ml medium) with Ad-APN-GFP (6000 virus particles / microliter medium) at a multiplicity of infection (MOI) of 100 (100 virus particles / one cell) . 72 hours after transfection, almost all islet cells expressed GFP ( figure 2 A), the transfection efficiency is close to 100%. At 24, 36, 48 and 72 hours after transfection, we further detected the expression of APN in islet cells by means of RT-PCR and WB. Sig...
Embodiment 3
[0031] Example 3 Aggregation of dispersed islet cells into functional islet-like clusters
[0032] In order to restore the function of islet cells similar to primary islet clusters, we washed the dispersed islet single cells with HBSS solution twice after virus transfection, and then inoculated them with 2×10 5 Concentration of cells / culture dish Inoculate in 35mm ultra-low adhesion culture dish and culture in RPMI-1640 medium for 4 days until the formation of islet-like clusters ( image 3 A), the cell viability of the islet-like group is higher than 95% ( image 3 B).
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com