Novel cell culture method, cell culture system and application of cell culture system

A culture system and in vitro cell culture technology, applied in biomaterials and biological fields, can solve complex and unfavorable problems, achieve the effects of protecting activity, promoting growth and differentiation, and benefiting growth and differentiation

Inactive Publication Date: 2019-09-24
兰溪市立顺生物有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The cells cultivated by two-dimensional cell culture technology gradually lose their original traits in the body in vitro, and are far from their natural growth state in the body in terms of shape, structure and function; The restriction of controlling factors and the interaction between in vivo and in vitro environments make it complicated, which is not conducive to exploring the specific mechanism

Method used

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  • Novel cell culture method, cell culture system and application of cell culture system
  • Novel cell culture method, cell culture system and application of cell culture system
  • Novel cell culture method, cell culture system and application of cell culture system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] A method for preparing a composite porous support, comprising:

[0063] The sodium chloride is sieved, and the particle size of the sieved sodium chloride is 283-300 μm, and then mixed with hydroxyapatite in a ratio of 1:0.03;

[0064] Pack the mixture of sodium chloride and hydroxyapatite tightly into a plastic syringe;

[0065] After mixing the liver cell activity factor HGF, collagen solution and silk fibroin solution at a ratio of 1:4, dissolve the double active ester polyethylene glycol in dimethyl sulfoxide, and then add it dropwise to the collagen and silk fibroin solution. In the mixed solution of plain protein, the concentration of polyethylene glycol of bisactive ester in the mixed solution is 0.02%, under certain pH value and temperature condition, stirring reaction;

[0066] Pour into the syringe, immediately press the plunger of the syringe, and place it at room temperature;

[0067] Remove the material from the syringe, soak it with ethanol, and then was...

Embodiment 2

[0069] A method for preparing a composite porous support, comprising:

[0070] The sodium chloride is sieved, and the particle size of the sieved sodium chloride is 283-300 μm, and then mixed with hydroxyapatite in a ratio of 1:0.03;

[0071] Pack the mixture of sodium chloride and hydroxyapatite tightly into a plastic syringe;

[0072] After mixing the liver cell activity factor HGF, collagen solution and silk fibroin solution at a ratio of 1:4, dissolve the double active ester polyethylene glycol in dimethyl sulfoxide, and then add it dropwise to the collagen and silk fibroin solution. In the mixed solution of plain protein, make the concentration of bisactive ester polyethylene glycol in the mixed solution be 0.02%, add catecholamine, make the concentration of catecholamine in the mixed solution be 0.04%, under certain pH value and temperature condition, stirring reaction;

[0073] Pour into the syringe, immediately press the plunger of the syringe, and place it at room t...

Embodiment 3

[0076] A method for preparing a composite porous support, comprising:

[0077] The sodium chloride is sieved, and the particle size of the sieved sodium chloride is 283-300 μm, and then mixed evenly with hydroxyapatite in a ratio of 1:0.02;

[0078] Pack the mixture of sodium chloride and hydroxyapatite tightly into a plastic syringe;

[0079] After mixing the liver cell activity factor HGF, collagen solution and silk fibroin solution at a ratio of 1:4, dissolve the double active ester polyethylene glycol in dimethyl sulfoxide, and then add it dropwise to the collagen and silk fibroin solution. In the mixed solution of plain protein, make the concentration of bisactive ester polyethylene glycol in the mixed solution be 0.03%, add catecholamine, make the concentration of catecholamine in the mixed solution be 0.05%, under certain pH value and temperature condition, stirring reaction;

[0080] Pour into the syringe, immediately press the plunger of the syringe, and place it at...

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Abstract

The invention provides a novel cell culture method, a cell culture system and application of the cell culture system, and belongs to the field of biological materials and biotechnology. The three-dimensional cell culture system provided by the invention is used for in-vitro cell culture and comprises a packed bed cell reactor, wherein the reactor comprises a composite porous scaffold, and the composite porous scaffold comprises collagen, silk fibroin and hydroxyapatite. The method for three-dimensional cell culturing by using the system comprises the following steps of: setting the three-dimensional cell culture system; supplying the cell reactor with cells and a cell culture solution; and circularly filling the reactor with the cell culture solution, wherein the cell inoculation mode is dynamic inoculation. The porosity of the composite porous scaffold of the cell culture system and the uniformity and permeability of the pore structure are good so as to facilitate the transportation and transmission of moisture, ions and other nutrient substances, to have higher sensitivity to temperature, to respond to the change of ambient temperature, and to control the subsequent release of growth factors.

Description

technical field [0001] The invention belongs to the field of biological materials and biotechnology, and specifically relates to a new cell culture method, a cell culture system and applications thereof. Background technique [0002] At present, in vitro cell-level research is largely done under two-dimensional culture conditions; in vivo, it is mainly done in experimental animal models. The cells cultivated by two-dimensional cell culture technology gradually lose their original traits in the body in vitro, and are far from their natural growth state in the body in terms of shape, structure and function; It is complicated by the restriction of controlling factors and the interaction between in vivo and in vitro environments, which is not conducive to exploring the specific mechanism. In view of the limitations of both, three-dimensional cell culture technology has been developed in recent years. The cells obtained by this culture method are significantly different from tho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12N5/00C12N5/071C12Q1/02
CPCC12M25/18C12N5/0062C12N5/0631C12N5/0671C12N2501/15C12N2503/02C12N2513/00C12N2533/18C12N2533/30C12N2533/50C12N2533/54G01N33/5008G01N2500/10
Inventor 车有泉郑宝华杨志军
Owner 兰溪市立顺生物有限公司
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