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.

Pending Publication Date: 2021-09-07
WEST CHINA HOSPITAL SICHUAN UNIV
View PDF9 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the severe loss of islet quantity and function shortly after transplantation limits its wide application.

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
  • Applications of adiponectin modified islet cells in improvement or enhancement of islet transplantation
  • Applications of adiponectin modified islet cells in improvement or enhancement of islet transplantation
  • Applications of adiponectin modified islet cells in improvement or enhancement of islet transplantation

Examples

Experimental program
Comparison scheme
Effect test

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).

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

The invention relates to the field of biological medicines, and in particular, to applications of adiponectin modification to enable islet cells to have antioxidant and / or anti-inflammatory characteristics and applications of adiponectin modification of the islet cells in improvement or enhancement of islet transplantation. The islet cells are subjected to adiponectin modification before islet transplantation, so that the islet cells overexpress adiponectin. The islet cells have the characteristics of oxidation resistance and inflammation resistance. Therefore, islet transplantation results are improved. During gene therapy, the islet cells are decomposed into single cells, so that the infection efficiency of viruses is greatly improved. After the gene therapy, the single cells are re-cultivated into functional cell clusters, so that pancreas islet functions are greatly reserved.

Description

technical field [0001] The present invention relates to the field of biomedicine, in particular to the application of adiponectin modification in the anti-oxidation and / or anti-inflammation of islet cells, and the application of adiponectin modification in islet cells to improve or enhance the effect of islet transplantation. Background technique [0002] Islet transplantation is considered to be a promising treatment for type 1 diabetes mellitus (T1DM). However, the severe loss of islet quantity and function shortly after transplantation limits its widespread application. Even in the absence of immune rejection such as allogeneic (autologous) islet transplantation, up to 60% of engrafted islets are rapidly destroyed, emphasizing the critical role of non-immune factors in regulating islet transplantation outcome. In the early stage after transplantation, the active oxygen free radicals and inflammatory response caused by the separation and purification of islets and ischemi...

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
Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/17A61K35/39A61P37/06A61P29/00A61P39/06C12N15/861C12N5/10
CPCC12N15/86C12N5/0676C07K14/47A61K48/0008A61K38/1709A61P37/06A61K35/39A61P29/00A61P39/06C12N2710/10043
Inventor 郑晓峰杜晓炯王成世陆燕蓉
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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