Chondrocytes isolation from hyaline cartilage by continuous monitoring method

Background: Articular cartilage has poor regenerative capacities. Numerous repair techniques are known, including implantation of autologous chondrocytes. Material and methods: From 18 rabbits pieces were harvested from femoral condyle. Minced was treated with 0.25% trypsin-EDTA. In the 1st group (n=9) digested 0.6% collagenase in 15 ml tubes by shaking incubator at 37°C, 5%CO2 . 2nd digestion performed 25cm2 cell culture flasks placed on lateral side, monitoring process under a microscope after 120 minutes. The isolated cells cultured to 80-90% confluence. chondrocytes identified using histochemical staining culturing for 16 days overconfluence. Results: Chondrocytes isolation lasted fixed 360 minutes, – 140±10 1stgroup 9.2x104 ±3.1x104 viability 85.36±16.41%, but 1.6x105 ±3.4x104 98.09±3.85%. mean period 15±2 days, 11±3 days. first passage obtained 1.2x106 ±4.3x105 2.92x106 ±3.6x105 secreted extracellular matrix stained specifically cartilaginous tissue. Conclusions: method used direct impact number cells, their viability, also upon during passage.