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Method for inducing mesenchymal stem cells (MSCs) into chondrocytes

A technology of chondrocytes and mesenchymal stem cells, applied in the field of stem cells, can solve the problems of long cycle and low efficiency of type II collagen expression

Active Publication Date: 2015-04-15
广东省医学医疗有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current methods that lead to the differentiation of MSCs into chondrocytes require a long cycle, usually 2-4 weeks, and the expression efficiency of type II collagen after induction is low

Method used

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  • Method for inducing mesenchymal stem cells (MSCs) into chondrocytes
  • Method for inducing mesenchymal stem cells (MSCs) into chondrocytes
  • Method for inducing mesenchymal stem cells (MSCs) into chondrocytes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] A method for inducing mesenchymal stem cells to become chondrocytes, comprising the steps of:

[0061] Step S11. Isolation and culture of umbilical cord MSCs:

[0062] Take the detached umbilical cord tissue of full-term normal and healthy puerpera from the operating table, fully wash the residual blood with PBS balance solution, remove the umbilical vein, artery and umbilical cord adventitia, peel off Wharton glue, and cut it into 1 mm 3 For small human tissue pieces, wash the blood cells with DMEM medium repeatedly, spread the tissue pieces in a 6cm-diameter petri dish with an interval of about 1cm between the tissue pieces, and place them in 37°C, 5% CO 2 Attach to the wall for 1h in the incubator. Add 1ml of DMEM containing 10% FBS to the Petri dish, carefully place it at 37°C, 5% CO 2 cultured in an incubator. Subculture is performed after cells have climbed out of the tissue and grown into a monolayer.

[0063] Step S12. Separation and cultivation of chondrocy...

Embodiment 2

[0078] A method for inducing mesenchymal stem cells to become chondrocytes, comprising the steps of:

[0079] Step S21. Isolation of MSCs and obtaining primary cultured MSCs:

[0080] Take the umbilical cord tissue of full-term normal and healthy puerpera from the operating table, fully wash the residual blood with PBS balance solution, remove the umbilical vein, artery and umbilical cord adventitia, peel off the Wharton glue, and cut it into 1 mm 3 For small human tissue pieces, wash the blood cells with DMEM medium repeatedly, spread the tissue pieces in a 6cm-diameter petri dish with an interval of about 1cm between the tissue pieces, and place them in 37°C, 5% CO 2 Attach to the wall for 1h in the incubator. Add 1ml of DMEM containing 10% FBS to the Petri dish, carefully place it at 37°C, 5% CO 2 Cultured in an incubator, and the primary cultured MSCs were obtained after the cells climbed out of the tissue;

[0081] Step S22. Isolation of chondrocytes and obtaining prim...

Embodiment 3

[0096] A method for inducing mesenchymal stem cells to become chondrocytes, comprising the steps of:

[0097] Step S31. Isolation of MSCs and obtaining primary cultured MSCs:

[0098] Take the umbilical cord tissue of full-term normal and healthy puerpera from the operating table, fully wash the residual blood with PBS balance solution, remove the umbilical vein, artery and umbilical cord adventitia, peel off the Wharton glue, and cut it into 1 mm 3 For small human tissue pieces, wash the blood cells with DMEM medium repeatedly, spread the tissue pieces in a 6cm-diameter petri dish with an interval of about 1cm between the tissue pieces, and place them in 37°C, 5% CO 2 Attach to the wall for 1h in the incubator. Add 1ml of DMEM containing 10% FBS to the Petri dish, carefully place it at 37°C, 5% CO 2 Cultured in an incubator, and the primary cultured MSCs were obtained after the cells climbed out of the tissue;

[0099] Step S32. Isolation of chondrocytes and obtaining prim...

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Abstract

The invention provides a method for inducing mesenchymal stem cells (MSCs) into chondrocytes. The method comprises the following steps of: obtaining MSCs and the chondrocytes; inoculating the chondrocytes into a chondrocyte culture solution to be cultured; inoculating MSCs in an inserted cell culture dish, placing the inserted cell culture dish in the chondrocyte culture solution again, and replacing the chondrocyte culture solution into a chondrocyte induced differentiation culture medium for co-culture. By adopting the induction method, MSCs and the chondrocytes can simulate an in-vivo environment to produce a synergy, so that induced MSCs are differentiated into chondrocyte factors to play a role of a synergistic effect for inducing factors with an extracellular matrix secreted by the chondrocytes, so as to obviously shorten the time for inducing MSCs to differentiate into chondrocytes, improve the proliferation rate of the chondrocytes, and simultaneously enhance the expression rate and the expression quantity of type II collagens which induce the differentiated cells and the secretion of glycosaminoglycan (GAG).

Description

technical field [0001] The invention belongs to the technical field of stem cells, and in particular relates to a method for inducing mesenchymal stem cells to become chondrocytes. Background technique [0002] At present, the traditional method of repairing cartilage tissue is to use autologous chondrocytes. In this method, the articular cartilage in the non-weight-bearing area must be excised under arthroscopy, and the chondrocytes in it must be expanded. The only advantage of this operation is that it has no antigenicity, but it causes Body damage is great. In addition, most of the cartilage tissue is matrix, and the number of chondrocytes is small and difficult to separate. The number of passages in vitro is limited, and generally cultured for 3 to 4 passages at most, so it is difficult to obtain a large number of cells. [0003] Currently, bone marrow mesenchymal stem cells are generally induced to induce chondrocytes by high-density aggregation. In this method, the a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077C12N5/0775
Inventor 赵文卓王涵刘韬
Owner 广东省医学医疗有限公司
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