[The possibility of isolation, culture and storage of articular cartilage cells].
Lesions of articular cartilage are a common problem and concern millions of people world-wide. A decrease in physical activity and pain symptoms among patients resulting from damage to articular cartilage have prompted research concerning new methods allowing cartilage regeneration. State-of-the-art treatment of articular damage depends very much on genetic engineering techniques. The aim of this paper was to determine the authors' own way of isolation, proliferation and storage of chondrocytes of articular cartilage. The material consisted of 30 rabbits, from which fragments of articular cartilage were taken. The study consisted of the following stages: isolation, chondrocyte proliferation, cell and matrix identification, storage and MTT tests. Matrix digestion was achieved using the following solutions: 0.1% type IA collagenase; 0.025% trypsin, a mixture of collagenase and trypsin. The greatest amount of cells were found after digestion of the basic matter of cartilage by 0.1% solution of type IA collagenase. When ascorbic acid was added to the medium, a 25% increase in cellularity was observed. A cumulation of procollagen mRNA was noted in the isolated cells. After about 21 days the isolated cells formed a multilayer structure, with the space between the cells filled with a substance that showed typical traits for cartilage matrix. Storing the isolated cells for less than 48 hours at room temperature gave a 90% survival rate. Most cells died after less than 12 hours when stored at 4 degrees C. The described method of chondrocyte isolation proved to be effective in preparing material for treatment of articular cartilage lesion.
Lecybył
R
R
Katedra i Zakład Biochemii i Biologii Molekularnej, Akademia Medyczna im. K. Marcinkowskiego w Poznaniu.
Trzeciak
T
T
Kruczyński
J
J
D002358:Cartilage, Articular / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division; D019902:Chondrocytes / Q000166:cytology*; D006801:Humans; D011347:Procollagen / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D014021:Tissue Preservation / Q000379:methods
D011347:Procollagen; D012333:RNA, Messenger