Preparation method and application of cross-linked hydrogel for muscle stem cell culture

A cross-linked hydrogel and stem cell culture technology, which is applied in the field of biological food materials, can solve the problems of poor mechanical properties and poor cell adhesion, achieve complete mechanical properties, improve biocompatibility, and facilitate adhesion of stem cells Effect

Active Publication Date: 2021-05-11
汕头得宝投资有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This form of hydrogel is biocompatible and more environmentally friendly, however, poor mechanical properties and poor cell adhesion are its major draw

Method used

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  • Preparation method and application of cross-linked hydrogel for muscle stem cell culture
  • Preparation method and application of cross-linked hydrogel for muscle stem cell culture
  • Preparation method and application of cross-linked hydrogel for muscle stem cell culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1: the preparation of chitosan / dextran / alginate / calcium ion double network cross-linked hydrogel

[0049] Add 1 g of sodium alginate (AR; 120 kDa; G / M ratio of 35 / 65) and 99 mL of deionized water to the beaker, and stir while adding until the sodium alginate dissolves to obtain a sodium alginate solution with a concentration of 1 wt%. Then add 0.1 g of calcium carbonate to the sodium alginate solution and stir until the calcium carbonate is uniformly dispersed to form a first gel material similar to mud. Then add 1 g of dextran (AR; 80-100 kDa) and 99 mL of deionized water to the beaker, stir until the dextran dissolves to obtain a 1 wt% dextran solution, add 2 g of chitosan (90.24% deacetylation; 230 kDa) into the dextran solution, and stir until the chitosan is evenly dispersed, forming a second gel mass similar to mud. Mix the two gel materials according to the ratio of the first gel material: the second gel material is 1:1, stir and mix evenly. The slur...

Embodiment 2

[0050] Example 2: Heparin-coated chitosan / dextran / alginate / calcium ion double network cross-linked hydrogel

[0051] Sodium heparin (0.5 g) and deionized water (100 mL) were added to the beaker to obtain a heparin solution (5.0 g / L). Prepared CS / Dex / Alg / Ca 2+ The hydrogel was soaked in PBS for 30 minutes, then soaked in heparin solution for 15 minutes, repeated three times, washed with PBS to remove unadsorbed heparin on the surface, and obtained CS / Dex / Alg / Ca coated with heparin by electrostatic adsorption. 2+ Hydrogels.

Embodiment 3

[0052] Example 3: Collagen coated chitosan / dextran / alginate / calcium ion double network cross-linked hydrogel

[0053] Collagen and deionized water were added to a beaker to obtain a collagen solution (20 wt%). The prepared CS / Dex / Alg / Ca coated with heparin 2+ The hydrogel was soaked in PBS for 30 minutes, then soaked in collagen solution for 15 minutes, repeated three times, washed with PBS to remove the collagen on the surface, through the interaction between collagen and heparin, a gel coated with collagen and heparin was obtained. CS / Dex / Alg / Ca 2+ Hydrogels. Subsequently, the hydrogel was freeze-dried in a vacuum freeze dryer (-80 °C).

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Abstract

The invention discloses a preparation method and application of cross-linked hydrogel for muscle stem cell culture, and belongs to the technical field of biological food materials. Chitosan, alginate, dextran and Ca<2+> are crosslinked through a physical crosslinking method to form double-network hydrogel with high mechanical strength, and then the hydrogel is coated with heparin and collagen through a dipping coating method, so that the hydrogel can immobilize growth factors and adhere to cells. Meanwhile, the extracted primary muscle stem cells are inoculated onto the hydrogel and cultured for 24 hours in a growth culture medium (79% of DMEM, 10% of FBS and 1% of double antibodies); and then the extracted primary muscle stem cells are cultured in an incubator with a differential medium (97% of DMEM, 2% of horse serum and 1% of double antibodies) for 7 days. By utilizing the hydrogel, the absorption of the muscle stem cells on nutrient substances can be enhanced, and the growth of the muscle stem cells is facilitated. The dual-network hydrogel has the potential to be used as a scaffold for muscle stem cell growth of stem cell culture meat.

Description

technical field [0001] The invention relates to a preparation method and application of a cross-linked hydrogel used for culturing muscle stem cells, in particular to a preparation method and application of a novel physical double-network cross-linked hydrogel, which belongs to the technical field of biological food materials. Background technique [0002] Stem cell cultured meat has the potential to reduce the environmental impact of meat products compared with traditional meat production (Environmental Science & Technology, 2015, 49, 11941-11949). Stem cell cultured meat has attracted increasing interest, and a variety of methods for the cultivation of meat from stem cells for food purposes have been proposed (Journal of Integrative Agriculture, 2015, 14, 222-233). However, the lack of scalable cell culture substrates (scaffolds) becomes a major challenge limiting stem cell culture. [0003] Artificial simulated extracellular matrix (ECM) hydrogels are used to reproduce t...

Claims

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Application Information

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IPC IPC(8): C08J3/075C08L5/08C08L5/02C08L5/04C08L5/10C08L89/00C08K3/26C12N5/077
CPCC08J3/075C12N5/0659C12N2533/74C12N2533/72C12N2533/70C12N2533/54C08J2305/08C08J2405/04C08J2405/02C08J2405/10C08J2489/00C08K2003/265C12N2537/10C12N2533/18C12N5/0658C08J3/24
Inventor 胡静尹健汪翔
Owner 汕头得宝投资有限公司
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