Cell inducing culture medium and method for inducing and culturing mesenchyma stem cells

A technology of cell induction and culture medium, applied in the field of tissue engineering, can solve the problems of low transformation rate and high cost of mesenchymal stem cells, and achieve the effects of strong activity, simple use process, and improved induction transformation rate

Inactive Publication Date: 2015-12-09
深圳华毓造血干细胞研究有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The technical problem to be solved by the present invention is that in the prior art, the co-cultivation of mesenchymal stem cells and chondrocytes to induce mesenchymal stem cells to transform into chondrocytes has defects such as high cost and low transformation rate of mesenchymal stem cells, Provided is a cell induction medium capable of individually inducing transformation of mesenchymal stem cells into chondrocytes and a method for inducing and culturing mesenchymal stem cells using the cell induction medium

Method used

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  • Cell inducing culture medium and method for inducing and culturing mesenchyma stem cells
  • Cell inducing culture medium and method for inducing and culturing mesenchyma stem cells
  • Cell inducing culture medium and method for inducing and culturing mesenchyma stem cells

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Embodiment 1

[0033] The method for further inducing and culturing chondrocytes of the present invention comprises the following steps:

[0034] 1. Obtain mesenchymal stem cells;

[0035] 2. Subculture the mesenchymal stem cells obtained in step 1;

[0036] 3. The mesenchymal stem cells subcultured in step 2 are induced and cultured using a cell induction medium.

[0037] In step 1, the process of obtaining mesenchymal stem cells is:

[0038]Air was injected into the marginal ear vein of New Zealand white rabbits to make them die. Use 75% alcohol to smear the limbs of the white rabbit to disinfect and reduce the pollution of rabbit hair. Cut off the femur and coelom of the rabbit limbs, remove the skin tissue and keep the muscle connective tissue at the joint, and cut off the muscle tissue in the remaining parts. Soak the treated tissue in 75% alcohol, take it out after 30 min, wash it with sterile PBS solution, and place it in an ultra-clean bench. Expose the joint by cutting away the...

Embodiment 2

[0043] The only difference from Example 1 is that in this example, the cell induction medium contains: the concentration of TGF-β is 10ng / ml, the concentration of dexamethasone is 10mmol / l, and the concentration of ascorbic acid is 70ug / ml, the concentration of sodium glycerophosphate is 10mmol / l, the concentration of ITS+Premix is ​​100ng / ml, the concentration of insulin is 6.25mg / ml, and the concentration of transferrin is 6.25mg / ml.

Embodiment 3

[0045] The only difference from Example 1 is that in this example, the cell induction medium contains: the concentration of TGF-β is 15ng / ml, the concentration of dexamethasone is 10mmol / l, and the concentration of ascorbic acid is 100ug / ml, the concentration of sodium glycerophosphate is 15mmol / l, the concentration of ITS+Premix is ​​150ng / ml, the concentration of insulin is 10mg / ml, the concentration of transferrin is 10mg / ml, and the concentration of glutamine is 10mmol / l .

[0046] In order to further verify that the cell induction medium for converting mesenchymal stem cells into chondrocytes provided by the present invention has significant beneficial effects, the present invention is verified by the following detection experiments:

[0047] 1. Detection object:

[0048] Control group: chondrocytes obtained by prior art;

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Abstract

The invention relates to a cell inducing culture medium, which comprises TGF-beta (Transforming Growth Factor-beta), dexamethasone, ascorbic acid, sodium glycerophosphate and ITS+Premix. The invention further provides a method using the cell inducing culture medium for inducing and culturing chondrocytes. The cell inducing culture medium can promote the mesenchyma stem cells to be converted into the chondrocytes, and the cell activity can be improved, so that by using the method for inducing and culturing the chondrocytes provided by the invention, the goal of singly inducting the mesenchyma stem cells to be converted into the chondrocytes can be achieved; the inducing conversion rate of the mesenchyma stem cells can be improved; and many chondrocytes can be obtained.

Description

technical field [0001] The invention relates to the technical field of tissue engineering, in particular to an induction medium and a method for inducing and culturing mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells are a kind of cells with multi-lineage differentiation potential, which can be directional induced into various cells such as islet-like cells and chondrocytes. Moreover, mesenchymal cells exist in various tissues of the human body, especially mesenchymal stem cells derived from umbilical cord, placenta, bone marrow, and adipose tissue. They have the advantages of a wide range of sources, easy collection, and no ethical issues. They can be induced to differentiate into cartilage on a large scale Cells provide a new technical solution for the clinical treatment of articular cartilage damage. However, at present, the co-cultivation of mesenchymal stem cells and chondrocytes is often used to induce the transformation of mesenchymal stem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 曾宪卓鲁菲
Owner 深圳华毓造血干细胞研究有限公司
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