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Culture method for enhancing differentiation from mesenchymal stem cells to chondrocytes

A chondrocyte and stem cell technology, applied in the field of stem cells, can solve the problems of severe dedifferentiation and low efficiency of chondrocytes, and achieve the effects of increasing the expression rate and expression amount, and increasing the proliferation rate.

Active Publication Date: 2018-12-18
丰泽康生物医药(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Such as adding autologous serum, etc., but the tendency of chondrocyte dedifferentiation is serious, and the efficiency after induction is low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A culture method for improving the differentiation of mesenchymal stem cells to chondrocytes, comprising the following steps:

[0031] (1) Extracting chondrocytes, inoculating the chondrocytes in cell culture medium for culturing; the extracted chondrocytes are primary cultured chondrocytes. The inoculum size of the chondrocytes was 0.6×10 4 ~0.8×10 4 piece / cm 2 . The chondrocytes were kept in the cell culture fluid for 1 day, and the cell culture fluid was the chondrocyte culture fluid.

[0032] (2) Extracting mesenchymal stem cells, seeding said mesenchymal stem cells in an insert cell culture dish; said extracted mesenchymal stem cells are primary cultured mesenchymal stem cells. The inoculum size of the mesenchymal stem cells was 3.5×10 4 ~4×10 4 piece / cm 2 .

[0033] (3) Place the insert-type cell culture dish described in step (2) together with the chondrocytes cultivated in step (1) in the cell induction differentiation medium to cultivate together; the c...

Embodiment 2

[0050] A culture method for improving the differentiation of mesenchymal stem cells to chondrocytes, comprising the following steps:

[0051] (1) Extracting chondrocytes, inoculating the chondrocytes in cell culture medium for culturing; the extracted chondrocytes are subcultured chondrocytes. The inoculum amount of the chondrocytes was 0.3×10 4 ~0.8×10 4 piece / cm 2 . The chondrocytes were in the cell culture fluid for 2 days, and the cell culture fluid was the chondrocyte culture fluid.

[0052] (2) extracting mesenchymal stem cells, seeding said mesenchymal stem cells in an insert cell culture dish; said extracted mesenchymal stem cells are primary cultured mesenchymal stem cells. The inoculum size of the mesenchymal stem cells was 3×10 4 ~4×10 4 piece / cm 2 .

[0053](3) Place the insert-type cell culture dish described in step (2) together with the chondrocytes cultivated in step (1) in the cell induction differentiation medium to cultivate together; the cultivation...

Embodiment 3

[0071] A culture method for improving the differentiation of mesenchymal stem cells to chondrocytes, comprising the following steps:

[0072] (1) Extracting chondrocytes, inoculating the chondrocytes in cell culture medium for culture; the extracted chondrocytes are a mixture of primary cultured or subcultured chondrocytes. The inoculum amount of the chondrocytes was 0.3×10 4 ~0.8×10 4 piece / cm 2 . The chondrocytes were kept in the cell culture fluid for 1 day, and the cell culture fluid was the chondrocyte culture fluid.

[0073] (2) Extracting mesenchymal stem cells, inoculating said mesenchymal stem cells in a plug-in cell culture dish; said extracted mesenchymal stem cells are one or both of primary cultured mesenchymal stem cells. kind of mix. The inoculum size of the mesenchymal stem cells was 3×10 4 ~5×10 4 piece / cm 2 .

[0074] (3) Place the insert-type cell culture dish described in step (2) together with the chondrocytes cultivated in step (1) in the cell in...

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Abstract

The invention belongs to the technical field of stem cells and particularly relates to a culture method for enhancing differentiation from mesenchymal stem cells to chondrocytes. The method includes:(1) extracting chondrocytes and inoculating the chondrocytes on a cell culture liquid for cultivation; (2) extracting the mesenchymal stem cells and inoculating the mesenchymal stem cells on an insertcell culture dish; (3) placing the insert cell culture dish and the chondrocytes, cultured in the step (1), together in a cell inducing differentiation culture medium for coculture, wherein the cellinducing differentiation culture medium containing human bone morphogenetic protein 2 and growth hormone HGH. In the invention, the chondrocytes and mesenchymal stem cells are cocultured on the inducing culture medium, wherein in-vivo environment is simulated under induction by the culture medium on the two cells, thus initiating synergistic effect, increasing proliferation rate of the chondrocytes and meanwhile simultaneously increasing the expression rate and quantity of collagen in the cells after the inducing differentiation.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and in particular relates to a culture method for improving the differentiation of mesenchymal stem cells into chondrocytes. 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. 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 the body The damage was great. In addition, most of the cartilage tissue is matrix, and the number of chondrocytes is small and difficult to separate. The nu...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775
CPCC12N5/0655C12N5/0662C12N2500/30C12N2500/32C12N2500/38C12N2501/105C12N2501/115C12N2501/148C12N2501/15C12N2501/155C12N2501/2301C12N2501/305C12N2501/33C12N2501/85C12N2501/999C12N2502/1317C12N2506/1346
Inventor 马亚东黄宗堂曹娜
Owner 丰泽康生物医药(深圳)有限公司
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