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
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[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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