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Preparation method of hydrogel for three-dimensional culture of tumor cells

A three-dimensional culture and tumor cell technology, applied in the field of high-throughput screening of anti-tumor drugs, can solve the problems of expensive animal gels, affecting the accuracy and objectivity of drug evaluation results, and limiting applications

Inactive Publication Date: 2013-01-30
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because animal gel contains many uncertain components and the component differences between batches affect the accuracy and objectivity of drug evaluation results; on the other hand, animal gel is expensive, which limits its wide application. application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] First, place 0.0975g of polymethylvinyl ether maleic anhydride in 10 mL of twice distilled water, heat at 90°C for two hours to obtain an aqueous solution of polymethylvinyl ether maleic acid; then add 0.0156g of crosslinking agent polyethylene Diol PEG (molecular weight: 10,000) was dissolved in 10 mL of double distilled water; the above crosslinking agent was added to polymethyl vinyl ether maleic acid aqueous solution (the amount of carboxyl group n -COOH : the amount of hydroxyl n -OH = 400:1), most of the water was removed by rotary evaporation under heating conditions, and then placed in an oven for cross-linking at 80°C for 24 hours to obtain a cross-linked hydrogel solid. Dissolve the above-mentioned hydrogel solid in 10 mL of double distilled water, then add 1.2500 g of esterifying agent polyethylene glycol monomethyl ether (MeO-PEG-OH (1K) and mix evenly under stirring conditions, and then rotary evaporate under heating conditions Remove most of the water, a...

Embodiment 2

[0036] First, place 0.0975g of polymethylvinyl ether maleic anhydride in 10 mL of twice distilled water, heat at 90°C for two hours to obtain an aqueous solution of polymethylvinyl ether maleic acid; then add 0.01563g of crosslinking agent polyethylene Glycol PEG (molecular weight is 10,000) and 1.2500g esterifying agent polyethylene glycol monomethyl ether are dissolved in 10 mL twice distilled water; -COOH : the amount of hydroxyl n -OH = 400:1) and the aqueous solution of the esterifying agent are added to the polymethyl vinyl ether maleic acid aqueous solution, and most of the water is removed by rotary evaporation under heating conditions, and then placed in an oven for cross-linking at 80°C At 24 h, a cross-linked hydrogel solid was obtained. Then the above-mentioned gel was dissolved in twice distilled water to form a 4wt% solution, and under stirring conditions, a saturated aqueous solution of sodium bicarbonate was added dropwise to the above-mentioned hydrogel solu...

Embodiment 3

[0038] First, place 0.7800g polymethyl vinyl ether maleic anhydride in 20 mL double distilled water, heat 90 ooC and hydrolyze for two hours to obtain polymethyl vinyl ether maleic acid aqueous solution; Diol PEG and poly(lactic-co-glycolic acid) triblock copolymer PLGA 1300 -PEG 1500 -PLGA 1300 (molecular weight 4100) was dissolved in 10 mL double distilled water to obtain an aqueous solution; the aqueous solution of the above crosslinking agent was added to the aqueous solution of polymethyl vinyl ether maleic acid (the amount of carboxyl group n -COOH : the amount of hydroxyl n -OH = 800:1), most of the water was removed by rotary evaporation under heating conditions, and then placed in an oven for cross-linking at 80°C for 24 hours to obtain a cross-linked hydrogel solid. Weigh 0.2000g of the hydrogel solid and dissolve it in 4.8000g of twice distilled water. Under stirring conditions, add a saturated aqueous solution of sodium bicarbonate dropwise to the above-mention...

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Abstract

The invention discloses a preparation method of a hydrogel for the three-dimensional culture of tumor cells. A methyl vinyl ether / maleic acid copolymer which is a polymethyl vinyl ether / maleic anhydride hydrolysate is cross-linked by a cross-linking agent which is polyethylene glycol, a poly(lactide-co-glycolide)-polyethylene glycol-poly(lactide-co-glycolide) triblock copolymer, a poly-epsilon-caprolactone lactide-polyethylene glycol-poly-epsilon-caprolactone lactide triblock copolymer or glucan; the proportion of a carboxyl group in the methyl vinyl ether / maleic acid copolymer and the proportion of a hydroxy group in the cross-linking agent are adjusted to obtain a gel of different cross-linking densities, superfluous carboxyl groups in the gel after the cross-linking are esterified by polyethylene glycol monomethyl ether; the obtained gel is processed to prepare a 5% aqueous solution; and the aqueous solution is neutralized with a weak alkali such as a saturated aqueous solution of sodium bicarbonate to the pH value of 7.0 in order to prepare the hydrogel which can be used as a tumor cell three-dimensional culture medium.

Description

technical field [0001] The invention relates to the field of preparation of medical materials, in particular to a method for preparing a hydrogel applicable to three-dimensional culture of tumor cells and high-throughput screening of antitumor drugs. [0002] Background technique [0003] Traditional two-dimensional tumor cell culture (2D culture) is widely used in tumor cell culture due to its simplicity, ease of operation, and low cost. However, due to the large difference between 2D culture and the real environment in vivo, such as tumor cells growing in a plane, a three-dimensional environment cannot be constructed, and the interaction between cells and cells or between cells and the matrix cannot be truly reflected. The in vitro three-dimensional cell culture (3D culture) microenvironment is closer to the cell growth environment in vivo, so 3D culture has been widely used in the biomedical field since the beginning of the 21st century. Applying 3D culture technolo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/075C08J3/24C08L35/08C12N5/09
Inventor 张天柱朱长皓顾宁陈峻崧窦骏
Owner SOUTHEAST UNIV
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