Method for culturing organoid by using elastin hydrogel

A technology of elastin and hydrogel, applied in the biological field, can solve the problems of intramolecular cross-linking, large molecular weight, and many reaction steps, and achieve improved cell adhesion, high efficiency of modified grafting, and high production efficiency Effect

Pending Publication Date: 2022-08-09
THE SEVENTH AFFILIATED HOSPITAL SUN YAT SEN UNIV SHENZHEN
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are still some problems in the existing literature on the preparation of hydrogels by modifying natural elastin (such as: Polymers, 2020, 12, 670), including many early reaction steps (two steps) and long time-consuming (the first step reaction 1 Hours, the second step reaction needs overnight), low efficiency (mercapto groups are prone to intramolecular crosslinking, which affects the subsequent grafting reaction; and mercapto groups can react with the double bonds at both ends of polyethylene glycol diac

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 for culturing organoid by using elastin hydrogel
  • Method for culturing organoid by using elastin hydrogel
  • Method for culturing organoid by using elastin hydrogel

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0040] Example 1

[0041] This embodiment provides a modification method of photocrosslinkable elastin, which is prepared according to the following steps: 0.05 g of soluble elastin is dissolved in 10 mL of 0.01 M phosphate physiological buffer at pH 8.0 to prepare an aqueous elastin solution. 0.01 g of methacrylate-polyethylene glycol-N-hydroxysuccinimide (polyethylene glycol polymerization degree of 4) was added to the above-mentioned elastin aqueous solution, and the reaction was stirred for 3 hours under normal temperature and nitrogen atmosphere. The reactants were dialyzed in a dialysis bag with a molecular weight cut-off of 3 kDa for 48 hours, and then the purified protein solution was lyophilized in a freeze dryer to obtain modified elastin.

Example Embodiment

[0042] Example 2

[0043] This embodiment provides a modification method for photocrosslinkable elastin, which is prepared according to the following steps: 0.2 g of soluble elastin is dissolved in 10 mL of 0.01 M phosphate physiological buffer at pH 8.0 to prepare an aqueous elastin solution. 0.1 g of methacrylate-polyethylene glycol-N-hydroxysuccinimide (polyethylene glycol polymerization degree of 20) was added to the above-mentioned elastin aqueous solution, and the reaction was stirred for 0.5 hour under normal temperature and nitrogen atmosphere. The reactants were dialyzed in a dialysis bag with a molecular weight cut-off of 3 kDa for 48 hours, and then the purified protein solution was lyophilized in a freeze dryer to obtain modified elastin.

Example Embodiment

[0044] Example 3

[0045] This example provides a modification method of photocrosslinkable elastin, which is prepared according to the following steps: 0.15 g of soluble elastin is dissolved in 10 mL of 0.01M phosphate physiological buffer at pH 8.3 to prepare an aqueous elastin solution. 0.08 g of methacrylate-polyethylene glycol-N-hydroxysuccinimide (polyethylene glycol polymerization degree of 10) was added to the above-mentioned elastin aqueous solution, and the reaction was stirred for 2 hours under normal temperature and nitrogen atmosphere. The reactants were dialyzed in a dialysis bag with a molecular weight cut-off of 3 kDa for 48 hours, and then the purified protein solution was lyophilized in a freeze dryer to obtain modified elastin.

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 discloses a method for culturing an organoid by using elastin hydrogel, which comprises the following steps: dissolving soluble elastin in deionized water or a buffer solution to prepare an elastin aqueous solution; the preparation method comprises the following steps: firstly, preparing an elastin, then adding acrylate-polyethylene glycol-N-hydroxysuccinimide or methacrylate-polyethylene glycol-N-hydroxysuccinimide to obtain modified elastin, and dialyzing, freezing and freeze-drying the modified elastin in deionized water to obtain dry modified elastin; dissolving the dry-state modified elastin in a phosphate physiological buffer solution, adding a photoinitiator, mixing with an organoid cell mass, and irradiating under a light source with a certain wavelength to obtain an elastin hydrogel/cell compound; and culturing in an organoid culture medium, and amplifying the cells in the hydrogel to obtain the organoid. The method disclosed by the invention is simple in process, rapid in reaction and high in elastin content, and the cultured organoid is stable in quality.

Description

technical field [0001] The invention belongs to the biological field, and in particular relates to a method for culturing organoids by utilizing elastin hydrogel. Background technique [0002] Organoids are multicellular tissues that reproduce in vivo tissue (partial) structure and function in three dimensions, formed by the self-assembly of stem cells or specific progenitor cells (mainly epithelial cells) in vivo in vitro. Thus, organoids are powerful models for studying tissue and organ development, wound repair, lesions, tumor tissue initiation, progression, metastasis, drug resistance, targeted therapy, and immunotherapy. Organoid culture currently relies primarily on soluble basement membrane extracts from mouse sarcomas, such as commercially available Matrigel. However, Matrigel has the problem of unclear composition, which affects the quality of organoid culture. It is a complex composed of a variety of proteins (collagen, laminin, etc.), polysaccharides and growth ...

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): C12N5/071C12N5/09C08G81/00
CPCC12N5/0679C12N5/0693C08G81/00C12N2533/50C12N2533/30C12N2535/00C12N2537/00C12N2529/10
Inventor 严乐平何裕隆张常华徐云升何勇陈伟万欢欢
Owner THE SEVENTH AFFILIATED HOSPITAL SUN YAT SEN UNIV SHENZHEN
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