Method for culturing organoid by using elastin hydrogel

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 ...

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 diacrylate simultaneously, causing grafting to The polyethylene glycol diacrylate on the elastin loses the photocrosslinking ability); and the molecular weight of the polyethylene glycol diacrylate used in the reaction is large, so that the protein content in the final product is only about 60%, and the polyethylene glycol content is high It may weaken the cell adhesion of the hydrogel; in addition, the grafting rate in this reaction is only about 60% (measure the change of primary amino group content before and after modification)

Images