Preparation method of two-ion starch-based stem cell expansion hydrogel and stem cell expansion and collection method

A stem cell and hydrogel technology, applied in the field of biomedical engineering, can solve the problems of non-specific protein interference, low amplification efficiency, inability to achieve large-scale expansion of stem cells, and difficulty in maintaining "stemness" to reduce biocompatibility Decrease, increase proliferative activity, reduce adhesion effect

Active Publication Date: 2018-04-20
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In clinical treatment and tissue engineering applications, the demand is relatively large, generally at 10 9 -10 11 Among them, the traditional two-dimensional culture technology cannot realize the large-scale expansion of stem cells due to the low expansion efficiency. In recent years, the developed 3D culture technology based on microcarriers has achieved the large-scale expansion of cells
However, there are still some problems that need to be solved. For example, it is easy to be interfered by non-specific proteins during the culture process, which makes it difficult to maintain the "stemness" of the cells during the expansion process, especially for some cells with self-differentiation ability (such as: Brown adipose stem cells with myocardial self-differentiation ability), this phenomenon is more obvious

Method used

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  • Preparation method of two-ion starch-based stem cell expansion hydrogel and stem cell expansion and collection method
  • Preparation method of two-ion starch-based stem cell expansion hydrogel and stem cell expansion and collection method
  • Preparation method of two-ion starch-based stem cell expansion hydrogel and stem cell expansion and collection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Preparation and Characterization of Methacrylate Modified Zwitterionic Starch (SB-ST-MA)

example 1

[0043] Example 1: 5g of starch was added to 20mL of water, 20mL of NaOH aqueous solution with a concentration of 25w / v / % was added dropwise, the temperature was raised to 40°C, and the reaction was carried out for 3 hours. Dissolve 2.5g of DCAPS in 250mL of water, add to the above reaction solution, heat to 80°C, react for 4 hours, cool to room temperature, and neutralize to neutral with glacial acetic acid. Then add 300mL of water, precipitate with 300mL of methanol, and dry under vacuum at room temperature to obtain diionic starch.

[0044] Fully dissolve 1.38g of zwitterionic starch and 4.0g of sodium chloride in 60mL of DMSO at 130°C, and keep stirring until the solution is cooled to 50°C. After adding 0.35 g of DMAP and continuing to stir for 30 minutes under nitrogen protection, 2.10 g of GMA was added to react for 36 hours. Pour the reaction mixture into a large amount of methanol to precipitate a tan powdery solid, filter it with suction, wash the crude product three ...

example 2

[0045] Example 2: Add 5g of starch to 5mL of water, add 5mL of NaOH aqueous solution with a concentration of 25w / v / % dropwise, raise the temperature to 30°C, and react for 0.5 hours. Dissolve 150g of DCAPS in 1500mL of water, add to the above reaction solution, heat to 50°C, react for 4 hours, cool to room temperature, and neutralize to neutral with glacial acetic acid. Then add 1500mL of water, precipitate with 7.5L of methanol, and dry under vacuum at room temperature to obtain diionic starch.

[0046] 3 g of zwitterionic starch and 1.38 g of sodium chloride were fully dissolved in 60 mL of DMSO at 130 °C, and the stirring was continued until the solution was cooled to 50 °C. After adding 2.07g DMAP under nitrogen protection and continuing to stir for 60 minutes, 0.69g GMA was added to react for 12 hours. Pour the reaction mixture into a large amount of methanol to precipitate a tan powdery solid, filter it with suction, wash the crude product three times with methanol and ...

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Abstract

The invention discloses a preparation method of a two-ion starch-based stem cell expansion hydrogel and a stem cell expansion and collection method. The hydrogel is prepared from metacrylic acid estermodified two-ion starch and double-ended sulfydryl modified polyethylene glycol in sulfydryl-alkene click means, and CGRGDS polypeptides are introduced in the preparation process, so that the affinity of the hydrogel for cells is improved. The mechanical strength of the hydrogel can be controlled within the range of 27.21 to 69.92 kPa through the content of a polymer. More importantly, the adhesion of nonspecific proteins in the cell expansion process can be effectively reduced. The hydrogel has high biocompatibility, the multiplication of brown fat stem cells can be supported, and the stem cell characteristic is maintained in the multiplication process. Solid technical support can be provided for tissue engineering and cell treatment strategy. The hydrogel is chemically obtained throughclicking, the participation of an organic solvent is eliminated, the environment is protected, and the disadvantage that the support biocompatibility is reduced, which is caused in the synthetic process, is reduced.

Description

technical field [0001] The invention relates to a stem cell expansion hydrogel and a preparation method thereof based on a two-ion starch hydrogel stem cell expansion and collection method, belonging to the field of biomedical engineering. Background technique [0002] Stem cells are the original source of the human body and its various tissue cells, and have the characteristics of high self-renewal ability, high proliferation and multi-lineage differentiation potential, implantability and reconstruction ability. It has good application prospects in cell therapy and tissue engineering, and its large-scale expansion is the premise and basis of its clinical application. In clinical treatment and tissue engineering applications, the demand is relatively large, generally at 10 9 -10 11 Among them, the traditional two-dimensional culture technology cannot realize the large-scale expansion of stem cells due to the low expansion efficiency. In recent years, the developed 3D cultu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/075C08B31/00C12N5/0775C08L3/04C08L7/02
CPCC08B31/00C08J3/075C08J2303/04C08J2471/02C12N5/0667C12N2533/30
Inventor 姚芳莲董殿宇李俊杰
Owner TIANJIN UNIV
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