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Animal cell culture microcarrier and preparation method thereof

A cell culture, microcarrier technology, applied in animal cells, vertebrate cells, biochemical equipment and methods, etc., can solve problems, safety of exogenous viruses in biological products, etc., achieve improved use efficiency, good water solubility and surface-active, easy-to-operate effects

Active Publication Date: 2022-02-18
PINGHU YOUPU BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the animal collagen of some microcarriers contains animal-derived components such as pigs and cattle, which will bring the risk of exogenous viruses in biological products and cause safety issues

Method used

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  • Animal cell culture microcarrier and preparation method thereof
  • Animal cell culture microcarrier and preparation method thereof
  • Animal cell culture microcarrier and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0176] Preparation of microcarriers containing 2,3-epoxypropyltrimethylammonium chloride (GTA) and arginine (polyarginine) ligands.

[0177] (1) GTA cross-linked with dextran microspheres or dextran flakes

[0178] Take 100g of dextran and add it to a round bottom flask, add 1kg of pure water, stir well to make it swell, add 110g of Na 2 SO 4 , after stirring at 30°C for 1 hour, add 15ml of 1M sodium hydroxide solution into the round bottom flask, stir to dissolve, then raise the temperature to 55°C, add 50g of GTA to react for 8 hours. The product was washed three times with absolute ethanol, washed once with purified water, and filtered with suction to obtain a dextran-GTA gel.

[0179] (2) Allyl glyceryl ether activated dextran microspheres or flakes

[0180] Take 100 g of the dextran-GTA prepared in step (1) and put it into a round bottom flask, add 1 kg of pure water, add 70 g of sodium hydroxide, stir to dissolve, add allyl glyceryl ether, and react at 55 ° C for 12 h...

Embodiment 2

[0186] Preparation of microcarriers containing GTA and lysine (polylysine) ligands.

[0187] The operation steps are the same as in Example 1, and the arginine (polyarginine) in step 4) is replaced with lysine (polylysine).

Embodiment 3

[0189] Preparation of microcarriers containing arginine (polyarginine) ligands.

[0190] (1) Allyl glyceryl ether activated dextran microspheres or flakes

[0191] Take 100g of dextran and put it into a round bottom flask, add 1kg of purified water, stir well to make it swell, add 110g of Na 2 SO 4 , Add 70g of sodium hydroxide into the round bottom flask, stir to dissolve, add allyl glyceryl ether, and react at 55°C for 12 hours. After 12 hours, the activated gel was washed successively with 10 times the volume of purified water, ethanol and purified water.

[0192] (2) Brominated Allyl Glyceryl Ether

[0193] Take 1 L of the activated gel prepared in step (2) and put it into a round bottom flask, and add 250 mL of purified water and 30 g of anhydrous sodium acetate into the round bottom flask. After stirring well, add bromine water into the round bottom flask until the liquid in the bottle turns yellow. After bromination for 10 minutes, sodium formate was added to termi...

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Abstract

The invention provides an animal cell culture microcarrier. The animal cell culture microcarrier provided by the invention is obtained by cross-linking different ligands on the surface of a treated matrix by using a cross-linking activator, so use safety is high, and the adherence speed of cells is remarkably increased; the surface of the microcarrier has an animal gelatin protein structure due to compound ionic ligands, amino acids or polypeptides in the microcarrier, so a good adhesion growth environment is provided for cells, and safety is remarkably improved; meanwhile, a homology difference between fish gelatin protein of the microcarrier and human protein is large, so the risk of animal-derived viruses is avoided; a preparation process of the microcarrier is simple and easy to implement, and raw materials are low in price; the prepared microcarrier can be applied to the production of large-scale cells, viruses and recombinant cell products (such as interferon, enzyme, nucleic acid and hormone); and the microcarrier provided by the invention has great potential in the aspect of large-scale culture of animal cells.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to an animal cell culture microcarrier and a preparation method thereof. Background technique [0002] Microcarrier culture: Microcarriers use tiny particles as the carrier for cell attachment, which can provide a considerable attachment area. Because the carrier is small in size and light in specific gravity, the cells can be suspended in the culture medium under mild stirring. A cell culture technique that ultimately enables cells to propagate into a monolayer on the surface of a carrier. As a research and production technology suitable for large-scale animal cell culture, microcarrier culture is very important for the study of the structure, function and differentiation of animal cells and the production of many important biological materials (such as enzymes, hormones, antibodies, nucleic acids, viruses, vaccines and Viral vectors for gene therapy) are of great significance. [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0602C12N2531/00C12N2533/50C12N2533/54C12N2533/70C12N2535/00
Inventor 张洪任郁英曹源涛石海涛
Owner PINGHU YOUPU BIOTECH CO LTD