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Micro-carrier for cell culture and preparation method and application thereof

A cell culture and microcarrier technology, applied in general culture methods, microcarriers, biochemical equipment and methods, etc., to achieve the effect of simple and easy preparation process, non-toxic raw materials and low cost

Inactive Publication Date: 2016-05-11
冯玉萍 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The starch microcarrier prepared from starch has a microporous structure, a large surface area, and is rich in accessible active groups such as hydroxyl groups, so it can effectively adsorb free components and become a good raw material for cells to adhere to the wall. At present, Starch microcarriers have not been reported

Method used

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  • Micro-carrier for cell culture and preparation method and application thereof
  • Micro-carrier for cell culture and preparation method and application thereof
  • Micro-carrier for cell culture and preparation method and application thereof

Examples

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preparation example Construction

[0064] A kind of preparation method step of microcarrier for cell culture of the present invention is as follows:

[0065] (1) First prepare gelatinized starch alkali solution: dissolve starch in deionized water, heat slowly until gelatinized while stirring, then add 0.5-2mol / L NaOH solution to continue stirring and mixing, cool and stand to form a uniform aqueous solution , in the aqueous phase solution, the mass volume percentage of starch is 0.1% to 15%;

[0066] (2) Mixing oil-water phase to make balls: Mix and stir non-toxic high-density oil, isooctane, dispersant and a certain amount of emulsifier to form a uniform oil phase solution, then slowly add the water phase solution in step (1), Stir to form a uniform and stable mixed phase; the sum of the volume concentration of the emulsifier and dispersant in the oil phase is 0.1% to 5%, and the volume ratio of the water phase solution to the oil phase solution is 1:1 to 10;

[0067] (3) Cross-linking and solidification into...

Embodiment 1

[0077] A kind of preparation method step of microcarrier for cell culture of the present invention is as follows:

[0078] (1) First prepare gelatinized starch alkali solution: weigh 10g of soybean starch and dissolve it in 200mL of deionized water, slowly heat to 80°C for gelatinization while stirring, then add 20mL of 0.5mol / L NaOH solution, continue to stir and mix, cool and stand into a uniform aqueous solution;

[0079] (2) Mix oil-water phase to make balls: Measure 500mL of soybean oil, 500mL of isooctane, 15mL of Span-8015mL and 10mL of Tween-8010mL and mix and stir for 20 minutes to form a uniform solution, then slowly add 220ml of water phase solution and stir for 30 minutes to form a uniform and stable mixed phase;

[0080] (3) Cross-linking and solidification into balls: slowly add 20 mL of polypropylene glycol diglycidyl ether into the mixed phase at a stirring speed of 500 rpm, and leave to stand after 6 hours of cross-linking reaction at room temperature;

[00...

Embodiment 2

[0088] A kind of preparation method step of microcarrier for cell culture of the present invention is as follows:

[0089] (1) First prepare gelatinized starch alkali solution: weigh 15g of potato starch and dissolve it in 500mL of deionized water, heat slowly to 80°C for gelatinization while stirring, then add 30mL of 1mol / L NaOH solution and continue to stir and mix, cool and stand still to form Homogeneous aqueous solution;

[0090] (2) Oil-water phase mixing balls: Measure 1000mL of engine oil, 1000mL of isooctane, Span-2050mL and Tween-8050mL, mix and stir for 20 minutes to form a uniform solution, slowly add 530ml of water phase solution, and stir for 30 minutes to form a uniform and stable mixture Mutually;

[0091](3) Cross-linking and solidification into balls: slowly add 60 mL of polyethylene glycol glycidyl ether into the mixed phase at a stirring speed of 700 rpm, and after 7 hours of cross-linking reaction at room temperature, let stand;

[0092] (4) Degreasing ...

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Abstract

The invention provides a preparation method of a micro-carrier for cell culture. The preparation method includes the following steps that 1, a gelatinized starch alkali solution is prepared firstly; 2, an oil phase and a water phase are mixed for pelleting; 3, crossing-linking and curing are conducted at normal temperature for pelletizing; 4, oil-out cleaning is conducted; 5, DEAE coupling is conducted; 6, washing is conducted. The invention further provides the micro-carrier prepared through the method and an application of the micro-carrier to cell culture. Common edible starch is adopted, the raw material is free of toxicity and wide in source, and the whole preparation process is simple and feasible; the method is controllable, cost is low, and industrial production is easy. By means of the micro-carrier, suspension culture can be conducted in a stirring state, the micro-carrier is suitable for large-scale suspension culture of multiple anchorage-dependent cells, and the micro-carrier can be widely applied to the field of biological pharmacy.

Description

technical field [0001] The invention relates to a microcarrier for cell culture and its preparation method and application. Background technique [0002] Animal cell bioreactor microcarrier culture technology is the leading technology for anchorage-dependent cell culture in the biopharmaceutical industry. Microcarriers are a class of non-toxic, non-rigid, uniform density, transparent small particles used in cell culture. The anchorage-dependent cells can be attached to the particle surface for suspension culture, thereby increasing the area of ​​cell attachment growth, which is conducive to large-scale culture and collection of cells. [0003] Since VanWezel first used DEAE-SephadexA50 as a microcarrier for adherent cell culture in 1967, there have been more than ten kinds of microcarriers for cell culture reported in research, including dextran microcarriers, polylysine liquid microcarriers, Macroporous gelatin microcarriers, cellulose microcarriers, chitosan microcarrier...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00C08J3/24
CPCC12N5/0075C08J3/24C08J2303/02C12N2531/00
Inventor 马忠仁李明生张磊靳冬武
Owner 冯玉萍
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