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Preparation method of surface for cell adherent growth under serum-free condition

A serum-free and cell-free technology, applied in biochemical equipment and methods, cell culture supports/coatings, microorganisms, etc., can solve problems such as reducing mass production costs and coating foaming, and improve shelf life and uniformity High, uniform monomer concentration distribution effect

Pending Publication Date: 2022-03-01
武汉中科先进材料科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In view of this, the present invention provides a method for preparing the surface of cells attached to the wall without serum, which solves the problem that the unevenness of the super-hydrophilic coating on the surface easily leads to the phenomenon of foaming during the curing process of the coating under the action of plasma. Good biocompatibility, meet the needs of cell adherent culture under serum-free conditions and reduce the cost of large-scale production

Method used

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  • Preparation method of surface for cell adherent growth under serum-free condition
  • Preparation method of surface for cell adherent growth under serum-free condition
  • Preparation method of surface for cell adherent growth under serum-free condition

Examples

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Effect test

Embodiment 1

[0027] (1) Plasma cleaning treatment: After placing the culture vessel in the plasma cavity, vacuumize to a vacuum degree of 5Pa. Under the conditions of plasma power 50W, radio frequency 13.56MHz, and air flow 50sccm, the polystyrene substrate was subjected to plasma discharge treatment for 3 minutes. This step can remove the release agent and hydrocarbons on the surface of the culture vessel, roughen the surface and introduce oxygen-containing functional groups to improve the hydrophilicity of the surface.

[0028] (2) Coating the mixed solution of acrylic acid and L-proline by ultrasonic atomization process: the culture vessel treated in step (1) was taken out of the plasma chamber and placed in an ultrasonic atomization sprayer. Mix acrylic acid aqueous solution with a concentration of 30% and L-proline with a purity of 99% at a mass ratio of 2:1, and the mixed solution is uniformly coated on the substrate with a spray flow rate of 50mL / min after ultrasonic atomization. m...

Embodiment 2

[0035](1) Plasma cleaning treatment: After placing the culture vessel in the plasma cavity, vacuumize to a vacuum degree of 15Pa. Under the conditions of plasma power 80W, radio frequency 13.56MHz, and air flow 50sccm, the polystyrene substrate was subjected to plasma discharge treatment for 6 minutes. This step can remove the release agent and hydrocarbons on the surface of the culture vessel, roughen the surface and introduce oxygen-containing functional groups to improve the hydrophilicity of the surface.

[0036] (2) Coating the mixed solution of acrylic acid and L-proline by ultrasonic atomization process: the culture vessel treated in step (1) was taken out of the plasma chamber and placed in an ultrasonic atomization sprayer. Mix acrylic acid aqueous solution with a concentration of 20% and L-proline with a purity of 99% at a mass ratio of 4:1, and the mixed solution is sprayed at a flow rate of 50mL / min and evenly coated on the substrate after ultrasonic atomization. ...

Embodiment 3

[0039] (1) Plasma cleaning treatment: After placing the culture vessel in the plasma cavity, vacuumize to a vacuum degree of 8Pa. At the beginning, under the conditions of plasma power 100W, radio frequency 13.56MHz, and air flow 10sccm, the polystyrene substrate was subjected to plasma discharge treatment for 10 minutes. This step can remove the release agent and hydrocarbons on the surface of the culture vessel, roughen the surface and introduce oxygen-containing functional groups to improve the hydrophilicity of the surface.

[0040] (2) Coating the mixed solution of acrylic acid and L-proline by ultrasonic atomization process: the culture vessel treated in step (1) was taken out of the plasma chamber and placed in an ultrasonic atomization sprayer. Mix acrylic acid aqueous solution with a concentration of 50% and L-proline with a purity of 99% at a mass ratio of 5:1. The mixed solution is sprayed at a flow rate of 30mL / min and evenly coated on the substrate after ultrasoni...

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Abstract

The invention relates to the technical field of surface treatment, and provides a preparation method of a surface for serum-free cell adherent growth. The polyacrylic acid-proline composite coating is obtained by coating the surface of a culture dish through ultrasonic atomization and performing plasma polymerization, the water contact angle is smaller than 10 degrees, proline plays a role in increasing the grafting amount of an acrylic monomer, the hydrophilic performance is more excellent, the biocompatibility is good, and the requirement of serum-free cell adherent culture can be met. The polyacrylic acid-proline composite coating is firmly combined with the surface, the effective time of the hydrophilic performance is long and can reach more than one year, and the shelf life of the culture vessel can be remarkably prolonged. Besides, an ultrasonic atomization spraying machine is adopted for coating the monomers, the concentration distribution of the monomers on the surface of the culture vessel is uniform, and the polyacrylic acid-proline composite coating synthesized through the plasma polymerization reaction is high in uniformity and does not blister in the curing process under the action of plasma.

Description

technical field [0001] The invention relates to the technical field of surface treatment, in particular to a method for preparing a surface for cell adherent growth without serum. Background technique [0002] As an important and commonly used technique in cell biology research methods, cell culture refers to the growth of cells under in vitro conditions, and the cells no longer form tissues during the culture process. According to the cell type, cell culture can be divided into suspension cell culture and adherent cell culture. For adherent cell culture, it is necessary to have a certain degree of hydrophilicity on the culture surface. However, the commonly used culture vessel material, polystyrene, is a hydrophobic material, so it is necessary to modify the surface of the substrate to achieve the attached growth of cells. There are two common surface treatment methods at present, one is to coat the culture surface with poly-D-lysine or collagen, and the other is to use s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00
CPCC12N5/00C12N2533/30
Inventor 康翼鸿喻学锋高明杨帆
Owner 武汉中科先进材料科技有限公司
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