Gel composition, biological scaffold gel as well as preparation method and application of biological scaffold gel
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A gel composition and bioscaffold technology, applied in the field of organoid culture, can solve the problems of increasing the difficulty of dissolving and recovering cells from the gel, and the culture effect is not good, and achieve the advantages of being conducive to cell survival and proliferation, simple and diverse components, guaranteeing The effect of stability
Active Publication Date: 2022-03-22
ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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Collagen has strong biocompatibility, but due to the lack of other materials such as cytokines, its culture effect is still not as good as matrigel from biological sources. In addition, pure collagen components are tightly cross-linked, which increases the ability of the gel to dissolve and recover cells. difficulty
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Embodiment 1
[0043] The present embodiment provides a kind of bioscaffold gel, comprises according to final concentration: polyethylene glycol (PEG), 0.5g / ml; Human type I collagen, 0.1g / ml; Laminin, 10ng / ml; B27, 1×; N2, 1×; N-acetylcystine, 10μM; Nicotinamide, 50μM; R-spondin1 50ng / ml; EGF, 50ng / ml; Azaindole-1, 1ng / ml; in HBSS buffer at pH = 7.0. The preparation method of the bio-scaffold gel comprises: according to the final concentration of the bio-scaffold gel to form the ingredients, first mix B27, N2, N-acetylcystine, Nicotinamide, Wnt signaling pathway agonist, EGF, ROCK inhibitor, and RGD polypeptide with a buffer Dissolve, then add polyethylene glycol, collagen, laminin and mix well to obtain. The bioscaffold gel is stored at -20 to -80°C.
Embodiment 2
[0045] The present embodiment provides a kind of bioscaffold gel, comprises according to final concentration: polyethylene glycol (PEG), 0.4g / ml; Mouse type IV collagen, 0.3g / ml; Laminin, 50ng / ml; B27, 0.5×; N2, 0.5×; N-acetylcystine, 1μM; Nicotinamide, 10μM; R-spondin1 100ng / ml; EGF, 500ng / ml; Y27632, 100ng / ml; medium. The preparation method and storage method of the bioscaffold gel are the same as in Example 1.
Embodiment 3
[0047] The present embodiment provides a kind of bioscaffold gel, comprises according to final concentration: polyethylene glycol (PEG), 0.05g / ml; Human type IV collagen, 0.3g / ml; Laminin, 50ng / ml; B27, 5×; N2, 1×; N-acetylcystine, 50μM; Nicotinamide, 150μM; R-spondin1 10ng / ml; EGF, 10ng / ml; Azaindole-1, 5ng / ml; in DMEM / F12 medium. The preparation method and storage method of the bioscaffold gel are the same as in Example 1.
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Abstract
The invention provides a gel composition, biological scaffold gel as well as a preparation method and application of the biological scaffold gel. The gel composition is prepared from polyethylene glycol (PEG), collagen, laminin, RGD polypeptide, B27, N2, N-acetylcystine, Nicotinamide, a Wnt signaling pathway agonist, an epidermal growth factor and a ROCK inhibitor. The biological scaffold gel comprises the following components according to final concentration: 0.01-0.5 g/ml of PEG (Polyethylene Glycol); the content of collagen is 0.02 to 0.5 g/ml; the concentration of the laminin is 0.05 to 100 ng/ml; b27, 0.5 to 5 *; n2, 0.5 to 5 *; the concentration of the N-acetylcystine is 0.1 to 50 [mu] M; the concentration of the Nicotinamide is 10 to 200 [mu] M; the concentration of the Wnt signal channel agonist is 25 to 500 ng/ml; eGF (Epidermal Growth Factor) with the concentration of The concentration of the ROCK inhibitor is 0.5 ng/ml to 100 ng/ml; the concentration of the RGD polypeptide is 0.1 mg/ml to 1 mg/ml; all the components are dissolved in a buffer solution or a basic culture medium. According to the method, 3D culture of cells and tissues can be efficiently carried out, and formed 3D cultures such as cell clusters and organoids can simulate in-vivo three-dimensional tissue structures and gene expression characteristics.
Description
technical field [0001] The invention relates to the technical field of organoid culture, in particular to a gel composition, a biological scaffold gel and a preparation method and application thereof. Background technique [0002] Organoids are a three-dimensional cell culture technology that can mimic the tissue structure and gene expression of in vivo organs in vitro. Organoids are a variety of cells including stem cells that undergo self-differentiation and self-assembly in vitro to form three-dimensional aggregates. The cell types, ratios, and spatial structures and arrangements of cells are highly similar to those in vivo and can simulate in vivo Some functions of organs. Furthermore, gene expression in organoids is closer to in vivo than in traditional 2D cell culture. Therefore, the results of organoid-based bioactivity assays are more relevant in vivo. Therefore, organoids are currently widely used in biological research, drug research, medical research, and the t...
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