Porous microcryogel cell three-dimensional culture carrier, and preparation method and preparation system thereof

A three-dimensional culture and preparation system technology, applied in the field of porous micro-ice gel cell three-dimensional culture carrier and its preparation, can solve the problems of cell phenotype change, inability to simulate the three-dimensional growth environment of cells, and the limited space of two-dimensional culture

Pending Publication Date: 2017-07-25
TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are many disadvantages in the traditional two-dimensional cell culture method, mainly including: 1. Two-dimensional plane culture cannot simulate the three-dimensional growth environment of cells under physiological conditions, and long-term culture will lead to changes in cell phenotype, which will bring obstacles to biomedical research
2. Two-dimensional culture is limited by the space of the plane, and large-scale cell expansion cannot be achieved

Method used

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  • Porous microcryogel cell three-dimensional culture carrier, and preparation method and preparation system thereof
  • Porous microcryogel cell three-dimensional culture carrier, and preparation method and preparation system thereof
  • Porous microcryogel cell three-dimensional culture carrier, and preparation method and preparation system thereof

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preparation example Construction

[0044] According to one aspect of the present invention, the present invention proposes a method for preparing a porous microcryogel cell three-dimensional culture carrier. The method for preparing a porous microcryogel cell three-dimensional culture carrier according to an embodiment of the present invention includes: preparing a mixed organic phase; Aqueous phase solution is added to the phase and emulsified by stirring method, ultrasonic method or microchannel method to obtain a uniform water-in-oil emulsion system; the water-in-oil emulsion system is filtered to leave the carrier material to be cleaned; the carrier material The material is washed to obtain a three-dimensional culture carrier of porous micro-ice jelly cells, wherein the emulsification is carried out at a temperature not higher than zero degrees Celsius.

[0045] Therefore, by adopting this method, the homogeneous mass production of the porous microcryogel cell three-dimensional culture carrier can be realize...

Embodiment 1

[0098] Prepare and mix the organic phase and place it in the reaction kettle, add the aqueous phase solution into the organic phase under the stirring action of the stirring paddle, emulsify through stirring to form uniformly dispersed emulsion droplets, and obtain porous micro-ice gel after filtering and washing Three-dimensional cell culture carrier. Wherein, during the emulsification process, the low-temperature circulation device is used to provide circulating low-temperature ethanol, and the temperature in the entire reactor is controlled to be maintained at -20 degrees Celsius; and the rotation speed of the stirring blade is controlled to be 1500rmp / min.

[0099] The microscopic image of the porous microcryogel cell three-dimensional culture carrier prepared by this method and re-immersed in aqueous solution after the final freeze-drying is as follows: image 3 As shown in (A), it shows that a uniform spherical microcryogel carrier with a particle size range of 80-150 μm...

Embodiment 2

[0101] Prepare and mix the organic phase and put it in the reaction kettle, add the aqueous phase solution into the organic phase under the stirring action of the stirring paddle, emulsify through stirring to form uniformly dispersed emulsion droplets, filter and wash to obtain porous micro-cryogel Three-dimensional cell culture carrier. Wherein, in the emulsification process, the low-temperature circulation device is used to provide circulating low-temperature ethanol, and the temperature in the entire reactor is controlled to be maintained at -30 degrees Celsius; and the rotation speed of the stirring blade is controlled to be 1000rmp / min.

[0102] The microscope images of the microcryogel cell culture carrier prepared by this method re-immersed in aqueous solution after the final freeze-drying are as follows: image 3 As shown in (B), it shows that a uniform spherical microcryogel carrier with a particle size range of 150-250 μm can be prepared on a large scale; the SEM ima...

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Abstract

The invention discloses a porous microcryogel cell three-dimensional culture carrier, and a preparation method and a preparation system thereof. The preparation method comprises the following steps: preparing a mixed organic phase; adding an aqueous phase solution to the mixed organic phase, and carrying out stirring process, ultrasonic process or microfluid process emulsifying to prepare a uniform water-in-oil emulsion system; filtering the water-in-oil emulsion system to leave a carrier material to be cleaned; and cleaning the carrier material in order to obtain the porous microcryogel cell three-dimensional culture carrier, wherein the emulsification step is carried out at a temperature not higher than 0 DEG C. The preparation method can realize the homogeneous mass production of the porous microcryogel cell three-dimensional culture carrier, and the porous microcryogel cell three-dimensional culture carrier has a large porosity, keeps the cell activity at a high level, promotes cell functions, has high mechanical elasticity, and can well protect the cultured cells from being damaged by high mechanical shearing force in a bioreactor in order to improve the survival rate of the cells.

Description

technical field [0001] The invention belongs to the field of biological materials. Specifically, the invention relates to a porous microcryogel cell three-dimensional culture carrier and a preparation method and a preparation system thereof. Background technique [0002] With the development of life science technology, relevant scientific research institutions and production enterprises have put forward higher requirements for cell culture. There are many disadvantages in traditional two-dimensional cell culture methods, mainly including: 1. Two-dimensional plane culture cannot simulate the three-dimensional growth environment of cells under physiological conditions. Long-term culture will lead to changes in cell phenotype, which will bring obstacles to biomedical research. 2. Two-dimensional culture is limited by the space of the plane, and large-scale cell expansion cannot be achieved. [0003] Based on the above problems, the three-dimensional porous cell culture carrier...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00C12M1/00B01J13/06
CPCB01J13/06C12M99/00C12N5/0062
Inventor 杜亚楠刘伟吕丞
Owner TSINGHUA UNIV
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