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Porous microsphere particle for three-dimensional cell culture and preparation method thereof

A technology of porous microspheres and granules, applied in general culture methods, cell culture supports/coatings, biochemical equipment and methods, etc., can solve problems such as high cost, high energy consumption, and uneven feed ratio, and achieve operational Convenience and low energy consumption

Pending Publication Date: 2020-09-25
SUZHOU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The three-dimensional cell culture carriers obtained by traditional freeze-drying are all in the form of lumps, which cannot be applied to large-scale bioreactors for suspension culture, and due to the low heat transfer efficiency, the preparation and processing process consumes a lot of energy, which brings great difficulties to actual production. come at a greater cost
Using microfluidic emulsification technology or atomizing droplets into liquid nitrogen to freeze and then vacuum freeze-dry the three-dimensional cultured microspheres, the size of the carrier microspheres is not uniform, and the pore size is difficult to control, which affects the relationship between the subsequent cells and the microspheres. Uneven feeding ratio
At the same time, a large amount of toxic and harmful reagents such as organic solvents and cross-linking reagents will be used in the production process, which increases the risk of operation and causes environmental pollution. Moreover, the cleanliness of liquid nitrogen is low, the process is complicated, and it is difficult to produce on a large scale

Method used

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  • Porous microsphere particle for three-dimensional cell culture and preparation method thereof
  • Porous microsphere particle for three-dimensional cell culture and preparation method thereof
  • Porous microsphere particle for three-dimensional cell culture and preparation method thereof

Examples

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Effect test

Embodiment 1

[0055] Prepare porous chitosan / gelatin microsphere particles for three-dimensional cell culture as follows:

[0056] (1) Select chitosan and gelatin (the ratio of chitosan and gelatin is 1:2) as the carrier material, dissolve in 2v / v% acetic acid solution, stir magnetically at 50°C until completely dissolved, the stirring speed is 600rpm, Afterwards, a chitosan / gelatin solution with a solid content of 3 wt % was obtained by homogenizing for 60 min under a pressure of 600 bar through a high-pressure homogenizer.

[0057] (2) Pour the chitosan / gelatin precursor solution configured in step (1) into the liquid storage tank, connect the microfluidic atomizer through a catheter, and atomize the precursor solution into small uniform droplets, the droplet size Controlled at about 200μm, the atomization pressure is 1.5kg / cm 3 , the working vibration frequency of the atomizer is 12kHz, and the amplitude is 15Vpp.

[0058] (3) The droplets in step (2) are spray-freezed to obtain ice ba...

Embodiment 2

[0061] (1) Gelatin is selected as the carrier material, dissolved in pure water with a solid content of 5wt%, magnetically stirred at 50°C until completely dissolved, and the stirring speed is 600rpm, and a 5wt% gelatin aqueous solution is prepared.

[0062] (2) Import the gelatin aqueous solution prepared in step (1) into the liquid storage tank, connect the microfluidic atomizer through the catheter, and atomize the precursor liquid into fine uniform droplets, and the droplet size is controlled at about 200 μm. The atomization pressure is 1kg / cm3, the working vibration frequency of the atomizer is 12kHz, and the amplitude is 15Vpp.

[0063] (3) The droplets in step (2) are spray-freezed to obtain ice ball particles with uniform size, the tower wall temperature is set to -80°C, and the cold air flow rate is set to 250L / min.

[0064] (4) Transfer the ice ball particles collected in step (3) to a vacuum freeze dryer, and freeze and dry them under vacuum for 72 hours to obtain g...

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Abstract

The invention relates to the technical field of cell culture, in particular to a porous microsphere particle for three-dimensional cell culture and a preparation method thereof. The preparation methodcomprises the following steps: dissolving chitosan and / or gelatin in a solvent, uniformly mixing to obtain a precursor solution; spraying the precursor solution by adopting a microfluidic atomizer toobtain liquid drops; carrying out spray freezing on the liquid drops to obtain ice ball particles; and carrying out vacuum freeze drying on the ice ball particles to obtain porous microsphere particles. A microfluidic spray freezing tower is combined with a vacuum freeze drying technology; by regulating and adjusting the formula of the precursor solution and technical parameters such as the sizeof atomized liquid drops, the freezing temperature, and the like, the particle size and the pore diameter of the dried carrier particles can be controlled, the adhesion, proliferation, differentiationand other properties of cells on the microsphere particles are optimized, the uniform and controllable three-dimensional cell culture microsphere particle with a porous structure is prepared, and thepreparation method has the advantages of low energy consumption and convenient operation.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a porous microsphere particle for three-dimensional cell culture and a preparation method thereof. Background technique [0002] Obtaining high-activity, high-purity, and sufficient number of stem cells is the primary condition for advancing stem cell therapy. At present, most stem cells are cultivated in the plate-adherent culture method. On the one hand, the adhesion of cells on plate culture in vitro is different from that in vivo, which may easily cause dryness loss. On the other hand, plate culture provides limited space for stem cells and cannot be obtained. sufficient number of stem cells. [0003] Three-dimension Cell Culture (TDCC), as an emerging in vitro cell culture technology, can not only preserve the material basis of the cell microenvironment in vivo, but also reflect the intuitiveness and controllability of cell culture. It refers to the co-cultivation of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J9/28C08J3/12C08L5/08C08L89/00C12N5/00
CPCC08J9/28C08J3/122C08J9/0061C12N5/0062C12N5/0075C08J2389/00C08J2405/08C08J2201/0482C08J2201/0484C12N2531/00C12N2533/54C12N2533/72
Inventor 吴铎严珅张盛宇孙振华黄园园
Owner SUZHOU UNIV
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