Serum-free chondrocyte culture medium and preparation method thereof

A chondrocyte and culture medium technology, which is applied in the direction of cell culture active agents, bone/connective tissue cells, biochemical equipment and methods, etc., can solve the problems of expensive, unavoidable heterologous bodies, conflicts, etc., and achieve low cost, It is conducive to large-scale application and the effect of shortening the time

Inactive Publication Date: 2017-06-13
叶宗耀
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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 chondrocyte culture medium and preparation method thereof
  • Serum-free chondrocyte culture medium and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1, a kind of serum-free chondrocyte culture medium

[0031] The serum-free chondrocyte culture medium includes basal medium and additives added in the basal medium, and the components of the additives include: cytokine 24ng / mL, polylysine 7mg / mL in terms of final concentration , arachidonic acid 2 μM, vitamin C 10 μg / mL, guayulelide 5 μg / mL, butylenediamine 5 μM, insulin 7 μg / mL, linolenic acid 3 μM, flavonoids 14 μg / mL, transferrin 12 μg / mL, polyvinylpyrrolidone 20μg / mL, glucagon 8μg / mL and fibronectin 20ng / mL.

[0032] The basal medium is RMPI 1640 medium.

[0033] The cytokines are composed of serum spreading factor, transforming growth factor and epidermal growth factor in a weight ratio of 8:7:3.

[0034] Preparation:

[0035] S1 Add cytokines, polylysine, arachidonic acid, vitamin C, guayulelide, butylenediamine, insulin, linolenic acid, flavonoids, transferrin and fibronectin to the basal medium, stir After 20 minutes, polyvinylpyrrolidone and gluc...

Embodiment 2

[0037] Embodiment 2, a kind of serum-free chondrocyte culture medium

[0038] The serum-free chondrocyte culture medium includes basal medium and additives added in the basal medium, and the ingredients of the additives include: cytokine 32ng / mL, polylysine 10mg / mL in terms of final concentration , arachidonic acid 5 μM, vitamin C 15 μg / mL, guayulelide 8 μg / mL, butylenediamine 9 μM, insulin 9 μg / mL, linolenic acid 6 μM, flavonoids 18 μg / mL, transferrin 16 μg / mL, polyvinylpyrrolidone 26 μg / mL, glucagon 13 μg / mL and fibronectin 32 ng / mL.

[0039] The basal medium is IMEM medium.

[0040] The cytokines are composed of serum spreading factor, transforming growth factor and epidermal growth factor in a weight ratio of 10:3:1.

[0041] Preparation:

[0042] S1 Add cytokines, polylysine, arachidonic acid, vitamin C, guayulelide, butylenediamine, insulin, linolenic acid, flavonoids, transferrin and fibronectin to the basal medium, stir After 40 minutes, polyvinylpyrrolidone and gl...

Embodiment 3

[0044] Embodiment 3, a kind of serum-free chondrocyte culture medium

[0045] The serum-free chondrocyte culture medium includes basal medium and additives added in the basal medium, and the ingredients of the additives include: cytokine 28ng / mL, polylysine 9mg / mL in terms of final concentration , arachidonic acid 3 μM, vitamin C 12 μg / mL, guayulelide 6 μg / mL, butanediamine 7 μM, insulin 8 μg / mL, linolenic acid 5 μM, flavonoids 16 μg / mL, transferrin 14 μg / mL, polyvinylpyrrolidone 22μg / mL, glucagon 11μg / mL and fibronectin 25ng / mL.

[0046] The basal medium is IMEM medium.

[0047] The cytokines are composed of serum spreading factor, transforming growth factor and epidermal growth factor in a weight ratio of 9:5:2.

[0048] Preparation:

[0049] S1 Add cytokines, polylysine, arachidonic acid, vitamin C, guayulelide, butylenediamine, insulin, linolenic acid, flavonoids, transferrin and fibronectin to the basal medium, stir In 26 minutes, polyvinylpyrrolidone and glucagon wer...

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Abstract

The invention belongs to the technical field of cell culture mediums, and specifically relates to a serum-free chondrocyte culture medium and a preparation method thereof. The serum-free chondrocyte culture medium comprise a basic culture medium and an additive, wherein the additive is prepared from the following ingredients in final concentration: 24-32ng/mL of cell factor, 7-10mg/mL of polylysine, 2-5 microns of arachidonic acid, 10-15 microgram/mL of vitamin C, 5-8 microgram/mL of parthenolide, 5-9 microns of butanediamine, 7-9 microgram/mL of insulin, 3-6 microns of linoleic acid, 14-18 microgram/mL of flavone, 12-16 microgram/mL of transferrin, 20-26 microgram/mL of polyvinylpyrrolidone, 8-13 microgram/mL of glucagon, and 20-32 ng/mL of fibronectin. The serum-free chondrocyte culture medium provided by the invention does not contain serum, is low in cost and in favor of scale application.

Description

technical field [0001] The invention belongs to the technical field of cell culture medium, and in particular relates to a serum-free chondrocyte culture medium 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 key to be solved at present. [...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0655C12N2500/25C12N2500/30C12N2500/38C12N2500/50C12N2500/92C12N2501/20C12N2501/335C12N2501/998
Inventor 叶宗耀
Owner 叶宗耀
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