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Method for cltivating cartilage cells in knee joint of BALB/c chmice

A chondrocyte and culture method technology, applied in bone/connective tissue cells, tissue culture, animal cells, etc., to achieve the effect of low cost, favorable for cell culture, and high survival rate

Inactive Publication Date: 2004-10-27
刘健
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no report on other methods of culturing chondrocytes in animal knee joints

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1 The concrete steps of the inventive method:

[0017] 1. Main experimental materials

[0018] (1) Source of chondrocytes: As animal materials, 8-16 female BALB / c (h-2d) 6-8 week old mouse knee articular cartilage specimens can be used.

[0019] (2) Digestive solution: 0.15-0.2% w / w collagenase type II solution, (Sigma, U.S.A), sterilized by filtration.

[0020] (3) Culture medium: DMEM medium (Gibco, U.S.A.) medium can be selected. Typically 12-18% w / w neonatal calf serum is added to the culture medium.

[0021] (4) Screen: 100-200 mesh copper mesh.

[0022] 2. Operation steps

[0023] (1) Taking materials and separating cells

[0024] 1) Cartilage tissue slices were cut from the surface of the knee joint of BALB / c mice with a scalpel blade, collected in phosphate buffered saline (PBS), and washed repeatedly to remove blood on the surface of the cartilage slices and synovial tissue that may be mistaken. Fresh cartilage is milky white with light blue, t...

Embodiment 2

[0032] Embodiment 2 uses the culture method of rabbit knee articular chondrocytes to cultivate BALB / c mouse knee articular chondrocytes and the culture method comparison of BALB / c mouse knee articular chondrocytes of the present invention

[0033] Using the culture method of rabbit knee articular chondrocytes to culture mouse knee articular chondrocytes, the survival rate after one week is 0-20%, and the survival rate is relatively low;

[0034] With the chondrocytes prepared by the method for culturing BALB / c mouse knee chondrocytes of the present invention, sufficient and well-differentiated Balb / c mouse knee chondrocytes can be obtained, and the survival rate can reach 80% to 90% in one week. The survival rate is high.

[0035] It can be seen from the above specific implementation methods that the method of the present invention obtains sufficient and well-differentiated mouse chondrocytes, and the survival rate of the cultured mouse chondrocytes can reach 80% to 90% after ...

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PUM

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Abstract

A method for culturing the cartilage cells of BALB / c mouse knee joint is disclosed. Its advantage is high surviral rate (80-90%).

Description

technical field [0001] The invention provides a method for culturing chondrocytes in the knee joints of BALB / c mice, specifically, a method for culturing cells using the knee joints of BALB / c mice as the source of chondrocytes, and belongs to the field of biotechnology. Background technique [0002] Repairing the damage of articular cartilage is a major clinical problem. At present, the bioengineering technology of biomaterials and cell engineering provides an option for the biological repair of cartilage. The existing technology is only the knee chondrocytes of rabbits. One kind of production and cultivation method, the materials selected in the method are: DMEM medium, type II collagenase, newborn calf serum. The specific separation steps of rabbit knee chondrocytes are as follows: the knee cartilage was obtained from 4 newborn Japanese big-eared white rabbits, the bilateral knee joints were cut under aseptic conditions, washed with phosphate buffer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
Inventor 刘健
Owner 刘健
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