Experimental method for enzymolysis, digestion and acquisition in vitro of rabbit knee cartilage unit
A technology of articular cartilage and experimental methods, applied in the direction of bone/connective tissue cells, vertebrate cells, animal cells, etc., can solve problems such as failure to form hyaline cartilage tissue, decrease in viscoelasticity, and degeneration of chondrocyte function, and achieve the acquisition method Simple, repeatable effect
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[0016] Concrete steps of the present invention are as follows:
[0017] (1) Rabbit knee joints were collected under aseptic conditions. The knee joint was opened in an ultra-clean workbench, and the full-thickness cartilage of the femur and tibial plateau of the knee joint was scraped with a sharp knife. The articular cartilage was repeatedly cut into pieces with small scissors, rinsed with sterile D-Hanks solution, the supernatant was discarded, weighed, and placed in a sterile Erlenmeyer flask for later use.
[0018] (2) Add 0.3% dispase (a neutral lyase, Sigma, USA) and 0.2% type II collagenase (Sigma, USA) to the Erlenmeyer flask at the rate of 12 mL DMEM-F12 culture solution / g cartilage. Both enzymes were dissolved in sterile DMEM-F12 culture solution (HyClone, USA), filtered through a 0.22 μm disposable filter, and sterilized. Place the Erlenmeyer flask containing the culture solution at 37°C, 5% CO 2 On the magnetic stirrer in the incubator, stir at a slow speed, and...
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