Mechanical environment culture method for improving proliferation activity of chondrocytes and maintaining phenotype of chondrocytes

A chondrocyte, proliferation activity technology, applied in the direction of bone/connective tissue cells, biochemical equipment and methods, culture process, etc., can solve problems such as lack of biomechanical properties, and achieve clinical and practical needs. The effect of increasing the proliferative activity

Inactive Publication Date: 2019-01-11
WEIFANG MEDICAL UNIV
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  • Description
  • Claims
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Problems solved by technology

In cartilage tissue engineering, if the seed cells are expanded and cultured in vitro under normal conditions, the tissue engineered cartilage formed by the cells has been proven to l...

Method used

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  • Mechanical environment culture method for improving proliferation activity of chondrocytes and maintaining phenotype of chondrocytes
  • Mechanical environment culture method for improving proliferation activity of chondrocytes and maintaining phenotype of chondrocytes
  • Mechanical environment culture method for improving proliferation activity of chondrocytes and maintaining phenotype of chondrocytes

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Embodiment Construction

[0030] 1. Isolation, culture and identification of chondrocytes:

[0031] One-month-old New Zealand white rabbits were killed by air embolism in the ear veins, the skin of the limbs was peeled off, the knee joint was exposed, the attached muscles, periosteum, synovium and other tissues around the joint were scraped off, and the cartilage tissue of the articular surface was cut out, and the cartilage slices were cut into 5mm ×5mm small pieces, washed 2-3 times with PBS solution. Add 0.25% trypsin / EDTA, digest at 37 °C for 30 min, remove the supernatant, add 0.2% type II collagenase, digest at 37 °C for 4-6 h, and filter through a 200-mesh sieve Remove impurities, centrifuge at 1000 r / min for 5 min, remove the supernatant, add DMEM / F12 complete medium containing 15% fetal bovine serum to make cell suspension, inoculate in culture flask at a cell concentration of 1×109 cells / L The cells were cultured in a CO2 incubator with saturated humidity, 37°C, and 5% volume fraction, and t...

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Abstract

The invention discloses a mechanical environment culture method for improving the proliferation activity of chondrocytes and maintaining phenotype of chondrocytes. The invention comprises the following steps: step 1, carrying out separation, culture and identification of chondrocytes; 2, inoculating the chondrocytes into a culture apparatus, loading the chondrocytes during the culture process, andculturing the chondrocyte in a mechanical environment so that the chondrocytes are mechanically stimulated. Under culture in mechanical environment, the proliferation of chondrocytes is improved, andco-cultured cells is promoted to have a good cartilage phenotype, which can quickly and effectively provide excellent seed cells for cartilage tissue engineering, and solve the clinical and practicalneeds of tissue engineering cartilage seed cells.

Description

technical field [0001] The invention relates to a mechanical environment culture method for improving chondrocyte proliferation activity and maintaining cartilage phenotype. Background technique [0002] Articular cartilage defects caused by trauma and various diseases are very common in clinical practice. With the aging of the population in my country, the number of patients is increasing day by day, but the self-repair function of articular cartilage is very limited, so the repair and reconstruction of articular cartilage damage has always been a longing for people It is particularly important to solve the problem of articular cartilage repair and related research. Tissue engineering has important application prospects in human disease treatment, rehabilitation, and health care. Cartilage tissue engineering in tissue engineering has brought hope to the repair of articular cartilage defects. [0003] At present, the repair and reconstruction of cartilage at home and abroad...

Claims

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Application Information

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IPC IPC(8): C12N5/077
CPCC12N5/0655C12N2502/1358C12N2527/00
Inventor 王国辉谢永芳丛杨杨晶
Owner WEIFANG MEDICAL UNIV
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