Chondrocyte proliferation in a new culture system.
OBJECTIVE
This study has aimed to study different culture systems that might stimulate an increase in cell proliferation of normal and osteoarthritis chondrocytes from articular cartilage in rat model.


METHODS
Three culture systems using chondrocytes embedded in alginate beads were tested: chondrocytes cultured in Dulbecco's modified Eagle's medium (DMEM) as control, a co-culture system consisting of a monolayer of de-differentiated chondrocytes as a source of mitotic factors, and an enriched medium containing culture medium obtained from a monolayer of chondrocytes and DMEM. Normal and osteoarthritis chondrocytes were stained with 5-carboxyfluorescein diacetate succinimidyl ester and were cultured in each of the three systems. After 5 days of culture cell, proliferation was detected by flow cytometry. Chondrocyte phenotype was confirmed by collagen type II and MMP-3 expression. To determine possible molecules released into the medium by the cultured chondrocyte monolayer and which would probably be involved in cell proliferation, a study of mRNA and expression of transforming growth factor-beta1 (TGF-beta1), fibroblastic growth factor-2 (FGF-2), epidermal growth factor (EGF), platelet derived growth factor-A (PDGF-A) and insulin-like growth factor-1 (IGF-1) proteins was conducted.


CONCLUSIONS
Chondrocytes in the co-culture system or in enriched medium showed an increase in proliferation; only when osteoarthritis chondrocytes were cultured in enriched medium would they display a statistically significant increase in their proliferation rate and in their viability. When chondrocytes from the monolayer were analysed, differential mRNA expression of TGF-beta1 and IGF-1 was found during all passages, which suggests that these two growth factors might be involved in chondrocyte proliferation.
Gomez-Camarillo
M A
MA
Experimental Pathology Deparment, Center for Research and Advance Studies, Mexico City, Mexico.
Almonte-Becerril
M
M
Vasquez Tort
M
M
Tapia-Ramirez
J
J
Kouri Flores
J B
JB
D000464:Alginates; D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology / Q000473:pathology; D018929:Cell Culture Techniques / Q000379:methods*; D054337:Cell Dedifferentiation; D049109:Cell Proliferation / Q000187:drug effects; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism / Q000473:pathology; D018920:Coculture Techniques / Q000379:methods; D024043:Collagen Type II / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000378:metabolism; D008297:Male; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D008938:Mitosis; D010003:Osteoarthritis / Q000473:pathology; D010982:Platelet-Derived Growth Factor / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism
D000464:Alginates; D024043:Collagen Type II; D017077:Culture Media, Conditioned; D006603:Hexuronic Acids; D010982:Platelet-Derived Growth Factor; D053773:Transforming Growth Factor beta1; C075278:platelet-derived growth factor A; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor; D007334:Insulin-Like Growth Factor I; D020723:Glucuronic Acid; D019278:Matrix Metalloproteinase 3