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Artificial cell membrane coating microcarrier for cell culture and preparation method thereof

A cell culture and cell membrane technology, applied in general culture methods, microcarriers, biochemical equipment and methods, etc., to achieve the effect of low price, long-term maintenance of cell activity and function, and uniform coverage

Inactive Publication Date: 2018-12-21
苏州怡彼得生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Konjac glucomannan (KGM) is a natural polymer polysaccharide with good biocompatibility and non-toxicity, etc. KGM microspheres have high transparency, good mechanical properties and suitable density, which is a potential microcarrier matrix. There are many domestic research reports, and many literatures and patents have introduced the synthesis method, such as: CN201210523380.8, but there are few reports on the surface modification of KGM microspheres for animal cell culture

Method used

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  • Artificial cell membrane coating microcarrier for cell culture and preparation method thereof
  • Artificial cell membrane coating microcarrier for cell culture and preparation method thereof

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Effect test

Embodiment 1

[0026] A preparation method of artificial cell membrane coating microcarriers for cell culture, comprising the steps of:

[0027] 1) Add 100 grams of konjac glucomannan microspheres to 500 ml of petroleum ether, stir and wash, filter with a Buchner funnel, then add to 500 ethanol, stir and wash, filter and dry for later use;

[0028] 2) Dissolve 29.5 g of 2-methacryloyloxyethyl phosphorylcholine (MPC) and 42.6 g of n-butyl methacrylate in 350 ml of ethanol, add 0.4 g of azobisisobutyronitrile (AIBN), Then the reaction system was evacuated with a vacuum pump and then fed with high-purity nitrogen, and this was repeated three times to replace the gas in the reaction system with high-purity nitrogen, wrapped with aluminum foil to avoid light, and then stirred and refluxed for 24 hours. The high-performance liquid phase detection raw material MPC has been basically consumed, stop the reaction, pour it into acetone with vigorous stirring to precipitate after cooling, wash with acet...

Embodiment 2

[0031] A preparation method of artificial cell membrane coating microcarriers for cell culture, comprising the steps of:

[0032] 1) Add 100 grams of konjac glucomannan microspheres to 500 ml of petroleum ether, stir and wash, filter with a Buchner funnel, then add to 500 ethanol, stir and wash, filter and dry for later use;

[0033] 2) Dissolve 29.5 g of 2-methacryloyloxyethyl phosphorylcholine (MPC) and 28.4 g of n-butyl methacrylate in 260 ml of ethanol, add 0.3 g of azobisisobutyronitrile (AIBN), Then the reaction system was evacuated with a vacuum pump and then fed with high-purity nitrogen. Repeatedly, the gas in the reaction system was replaced with high-purity nitrogen. After wrapping it with aluminum foil to avoid light, it was stirred and refluxed for 24 hours. The high-performance liquid phase detection raw material MPC has basically Consumed, stopped the reaction, cooled and poured into vigorously stirred acetone to precipitate, filtered and washed with acetone thr...

Embodiment 3

[0036] A preparation method of artificial cell membrane coating microcarriers for cell culture, comprising the steps of:

[0037] 1) Add 100 grams of konjac glucomannan microspheres to 500 ml of petroleum ether, stir and wash, filter with a Buchner funnel, then add to 500 ethanol, stir and wash, filter and dry for later use;

[0038] 2) Dissolve 29.5 g of 2-methacryloyloxyethyl phosphorylcholine (MPC) and 42.6 g of n-butyl methacrylate in 350 ml of ethanol, add 0.4 g of azobisisobutyronitrile (AIBN), Then the reaction system was evacuated with a vacuum pump and then fed with high-purity nitrogen. Repeatedly, the gas in the reaction system was replaced with high-purity nitrogen. After wrapping it with aluminum foil to avoid light, it was stirred and refluxed for 24 hours. The high-performance liquid phase detection raw material MPC has basically Consumed, stopped the reaction, cooled and poured into vigorously stirred acetone to precipitate, filtered and washed with acetone thr...

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Abstract

The invention provides an artificial cell membrane coating microcarrier for cell culture and a preparation method thereof. The microcarrier provided by the invention takes konjac glucomannan microspheres as a matrix, and an artificial cell membrane coating microsphere is made by a biomimetic new material. The preparation method of the microcarrier comprises the following steps: respectively washing the matrix microspheres with petroleum ether and absolute ethyl alcohol, and drying for later use; dissolving 2-methacryloyloxyethyl phosphate choline (MPC) and n-butyl methacrylate in ethanol, polymerizing into an artificial cell membrane material under the effect of an initiator azodiisobutyronitrile (AIBN), diluting to 2 percent concentration for later use; adding the washed konjac glucomannan microspheres to an artificial cell membrane ethanol solution and soaking, and drying by a rotary evaporator in a vacuum, thus obtaining a microcarrier product. The artificial cell membrane materialcovers the surface of the microcarrier evenly and is connected firmly, has the advantages of avirulence, good cell adhesion, degradability, capability of easily eluting cells without adhesion, and thelike, the cost of the prepared microcarrier is low, the production process is simple, and the period is shorter, thus being beneficial for industrial production.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering of biological materials, in particular to an artificial cell membrane-coated microcarrier for cell culture and a preparation method thereof. Background technique [0002] Microcarrier culture technology is currently recognized as the most promising technology for large-scale culture of animal cells. Since Van Wezel first used microcarriers for animal cell culture in 1967, microcarrier culture technology has developed into the main method for large-scale culture of anchorage-dependent cells. Microcarriers have the advantages of large specific surface area, high utilization rate of medium, and simple cell collection process. They are widely used in the cultivation of various cells and the production of biological products such as vaccines, antibodies, and nucleic acids. They also have certain applications in the fields of tissue engineering and regenerative medicine. application prospe...

Claims

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Application Information

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IPC IPC(8): C12N5/00C08F220/18C08F230/02
CPCC08F220/18C12N5/0075C12N2531/00C08F220/1804C08F230/02
Inventor 谢同张建军
Owner 苏州怡彼得生物技术有限公司
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