Serum-free medium applicable to chondrocyte in vitro culture and preparation method of serum-free medium

A serum-free medium and in vitro culture technology, which is applied to cell culture active agents, bone/connective tissue cells, biochemical equipment and methods, etc., can solve problems such as conflicts, high prices, and inability to avoid heterologous bodies, and achieve shortening Time and cost are low, and it is conducive to the effect of large-scale application

Inactive Publication Date: 2017-03-22
JIANGXI YIXINTANG MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are certain disadvantages in the serum medium, which cannot avoid the heterologous body in the animal serum. If the cells cultivated in the animal serum medium are used, there will be rejection and conflict of the human heterologous body.
Although some media do not contain serum, they are expensive

Method used

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  • Serum-free medium applicable to chondrocyte in vitro culture and preparation method of serum-free medium
  • Serum-free medium applicable to chondrocyte in vitro culture and preparation method of serum-free medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1, a kind of serum-free medium suitable for chondrocyte culture in vitro

[0024] The serum-free medium suitable for in vitro culture of chondrocytes includes a basal medium and additives added to the basal medium, and the ingredients of the additives include: butanediamine 2 μM, serine protease inhibitory Protein 10 μg / mL, fibronectin 15 ng / mL, transferrin 25 μg / mL, insulin 5 μg / mL, transforming growth factor 1 ng / mL, epidermal growth factor 3 ng / mL, polylysine 2 mg / mL, arachidonic acid Acid 1 μM, linolenic acid 4 μM, serum spreading factor 4 ng / mL, vitamin C 15 μg / mL, guayulelide 10 μg / mL, flavone 20 μg / mL.

[0025] The basal medium is DMEM medium.

[0026] Preparation:

[0027] S1 Add butanediamine, serpin, fibronectin, transferrin, insulin, transforming growth factor, epidermal growth factor, polylysine, arachidonic acid, linolenic acid, serum spreading to the basal medium factor, vitamin C, guayulelide and flavonoids, mixed evenly to obtain a mixture;...

Embodiment 2

[0029] Embodiment 2, a kind of serum-free medium suitable for chondrocyte culture in vitro

[0030] The serum-free medium suitable for in vitro culture of chondrocytes includes a basal medium and additives added to the basal medium, and the ingredients of the additives include: butanediamine 3 μM, serine protease inhibitory Protein 20 μg / mL, fibronectin 25 ng / mL, transferrin 35 μg / mL, insulin 10 μg / mL, transforming growth factor 10 ng / mL, epidermal growth factor 15 ng / mL, polylysine 5 mg / mL, arachidonic acid Acid 3μM, linolenic acid 10μM, serum spreading factor 11ng / mL, vitamin C 25μg / mL, guayulelide 30μg / mL, flavone 32μg / mL.

[0031] The basal medium is F12 medium.

[0032] Preparation:

[0033] S1 Add butanediamine, serpin, fibronectin, transferrin, insulin, transforming growth factor, epidermal growth factor, polylysine, arachidonic acid, linolenic acid, serum spreading to the basal medium factor, vitamin C, guayulelide and flavonoids, mixed evenly to obtain a mixture; ...

Embodiment 3

[0035] Embodiment 3, a kind of serum-free medium suitable for chondrocyte culture in vitro

[0036]The serum-free medium suitable for in vitro culture of chondrocytes includes a basal medium and additives added to the basal medium, and the ingredients of the additives include: butanediamine 2.5 μM, serine protease Arrestin 16μg / mL, fibronectin 20ng / mL, transferrin 28μg / mL, insulin 8μg / mL, transforming growth factor 3ng / mL, epidermal growth factor 10ng / mL, polylysine 4mg / mL, peanut four enoic acid 2 μM, linolenic acid 6 μM, serum spreading factor 7 ng / mL, vitamin C 20 μg / mL, guayulelide 22 μg / mL, flavonoids 25 μg / mL.

[0037] The basal medium is F12 medium.

[0038] Preparation:

[0039] S1 Add butanediamine, serpin, fibronectin, transferrin, insulin, transforming growth factor, epidermal growth factor, polylysine, arachidonic acid, linolenic acid, serum spreading to the basal medium factor, vitamin C, guayulelide and flavonoids, mixed evenly to obtain a mixture;

[0040] S...

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Abstract

The invention belongs to the technical field of cell mediums, and in particular relates to a serum-free medium applicable to chondrocyte in vitro culture and a preparation method of the serum-free medium. The medium consists of a basal culture medium and additives, wherein the additives, in terms of final concentration, include the following components: 2-3 [mu]M of butanediamine, 10-20 [mu]g / mL of serine protease inhibitor protein, 15-25ng / mL of fibronectin, 25-35 [mu]g / mL of transferrin, 5-10 [mu]g / mL of insulin, 1-10ng / mL of transforming growth factors, 3-15ng / mL of epidermal growth factors, 2-5mg / mL of polylysine, 1-3 [mu]M of arachidonic acid, 4-10 [mu]M of linolenic acid, 4-11ng / mL of serum spread factors, 15-25 [mu]g / mL of vitamin C, 10-30 [mu]g / mL of parthenolide and 20-32 [mu]g / mL of flavone. The medium provided by the invention is relatively low in cost.

Description

technical field [0001] The invention belongs to the technical field of cell culture medium, and in particular relates to a serum-free culture medium suitable for culturing chondrocytes in vitro and a preparation method thereof. Background technique [0002] Articular cartilage is mainly avascular tissue composed of chondrocytes and extracellular matrix. It mainly relies on joint movement and extrusion to absorb nutrients. Therefore, the self-repair ability of cartilage after injury is relatively weak. one of the problems. In recent years, with the development of tissue engineering technology, autologous cartilage transplantation has been applied in the clinical treatment of cartilage injury at home and abroad. Clinical research has limited patient tissues, and because chondrocytes are terminally differentiated cells, their in vitro proliferation ability is limited and they are easy to dedifferentiate, so the cultivation and expansion of in vitro chondrocytes has become the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0655C12N2500/25C12N2500/36C12N2500/38C12N2500/46C12N2500/50C12N2500/90C12N2501/11C12N2501/148C12N2501/15C12N2501/734C12N2501/998C12N2501/999
Inventor 胡波钟勇财谢玉国
Owner JIANGXI YIXINTANG MEDICAL TECH
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