Dedifferentiated chondrocytes cultured in alginate beads: restoration of the differentiated phenotype and of the metabolic responses to interleukin-1beta.
Chondrocytes cultivated in monolayer rapidly divide and lose their morphological and biochemical characteristics, whereas they maintain their phenotype for long periods of time when they are cultivated in alginate beads. Because cartilage has a low cellularity and is difficult to obtain in large quantities, the number of available cells often becomes a limiting factor in studies of chondrocyte biology. Therefore, we explored the possibility of restoring the differentiated properties of chondrocytes by cultivating them in alginate beads after two multiplication passages in monolayer. This resulted in the reexpression of the two main markers of differentiated chondrocytes: Aggrecan and type II collagen gene expression was strongly reinduced from day 4 after alginate inclusion and paralleled protein expression. However, 2 weeks were necessary for total suppression of type I and III collagen synthesis, indicators of a modulated phenotype. Interleukin-1beta, a cytokine that is present in the synovial fluid of rheumatoid arthritis patients, induces many metabolic changes on the chondrocyte biology. Compared with cells in primary culture, the production of nitric oxide and 92-kDa gelatinase in response to interleukin-1beta was impaired in cells at passage 2 in monolayer but was fully recovered after their culture in alginate beads for 2 weeks. This suggests that the effects of interleukin-1beta on cartilage depend on the differentiation state of chondrocytes. This makes the culture in alginate beads a relevant model for the study of chondrocyte biology in the presence of interleukin-1beta and other mediators of cartilage destruction in rheumatoid arthritis and osteoarthrosis.
Lemare
F
F
Laboratoire de Pharmacologie Cellulaire de l'Ecole Pratique des Hautes Etudes, Centre de Recherches Biomédicales des Cordeliers, Paris, France.
Steimberg
N
N
Le Griel
C
C
Demignot
S
S
Adolphe
M
M
D053674:Aggrecans; D000423:Alcian Blue; D000464:Alginates; D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D019902:Chondrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics; D003094:Collagen / Q000096:biosynthesis / Q000235:genetics; D004396:Coloring Agents; D016326:Extracellular Matrix Proteins; D015870:Gene Expression / Q000502:physiology; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000494:pharmacology*; D037181:Lectins, C-Type; D008863:Microspheres; D010641:Phenotype; D011509:Proteoglycans / Q000096:biosynthesis / Q000235:genetics; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D013194:Staining and Labeling
D053674:Aggrecans; D000464:Alginates; D011508:Chondroitin Sulfate Proteoglycans; D004396:Coloring Agents; D016326:Extracellular Matrix Proteins; D007375:Interleukin-1; D037181:Lectins, C-Type; D011509:Proteoglycans; D012333:RNA, Messenger; D012313:RNA; D003094:Collagen; D000423:Alcian Blue