Fibroblast growth factor-2 in serum-free medium is a potent mitogen and reduces dedifferentiation of human ear chondrocytes in monolayer culture.
The loss of the differentiated phenotype (dedifferentiation) during the expansion culture of donor chondrocytes remains a large problem in cartilage tissue engineering. Dedifferentiated chondrocytes produce other matrix components and therefore the tissue produced will be of less suitable quality. Previously, the addition of fibroblast growth factor-2 (FGF2) to a serum-containing medium (SCM) during expansion culture was shown to have positive effects on the phenotype of articular chondrocytes. In the present study, we focused on a more defined, serum-free medium (SFM), to expand chondrocytes in monolayer culture for the purpose of cartilage tissue engineering. Adult human ear chondrocytes were expanded in serum-free medium supplemented with 100 ng/ml FGF2. Expansion culture in a conventional serum-containing medium (10% FCS) served as control. The cell yield during expansion culture in serum-free medium with FGF2 was significantly higher compared to serum-containing medium. In addition, chondrocytes expanded in the serum-free medium with FGF2 expressed a more differentiated phenotype at the end of monolayer culture, as indicated by higher gene expression ratios of collagen type II to collagen type I and aggrecan to versican. Also, a higher gene expression of Sox9 was found. Next, suspension in alginate and subsequent culture in vitro or subcutaneous implantation in nude mice was used to evaluate the capacity of the chondrocytes, expanded in either medium, to re-express the differentiated phenotype (redifferentiation) and to form cartilage. The observed beneficial effects of the serum-free medium with FGF2 on the chondrocyte phenotype at the end of monolayer culture were sustained on both transcriptional and extracellular level throughout both redifferentiation methods.
Department of Orthopaedics, Erasmus MC, Room Ee 1659b, PO Box 1738, 3000 DR Rotterdam, The Netherlands.
E W
EW
Mandl
H
H
Jahr
J L M
JL
Koevoet
J P T M
JP
van Leeuwen
H
H
Weinans
J A N
JA
Verhaar
G J V M
GJ
van Osch
D000328:Adult; D053674:Aggrecans; D000464:Alginates / Q000494:pharmacology; D000818:Animals; D002356:Cartilage / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000502:physiology; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism; D003094:Collagen / Q000378:metabolism; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D004423:Ear; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D020723:Glucuronic Acid / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D006609:High Mobility Group Proteins / Q000378:metabolism; D006801:Humans; D037181:Lectins, C-Type; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D008934:Mitogens / Q000494:pharmacology*; D010641:Phenotype; D011509:Proteoglycans / Q000378:metabolism; D055758:SOX9 Transcription Factor; D023822:Tissue Engineering; D014157:Transcription Factors / Q000378:metabolism; D053675:Versicans
C508320:Acan protein, mouse; D053674:Aggrecans; D000464:Alginates; D011508:Chondroitin Sulfate Proteoglycans; D016895:Culture Media, Serum-Free; D016326:Extracellular Matrix Proteins; D006603:Hexuronic Acids; D006609:High Mobility Group Proteins; D037181:Lectins, C-Type; D008934:Mitogens; D011509:Proteoglycans; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; C527933:Sox9 protein, mouse; D014157:Transcription Factors; C508322:VCAN protein, human; C508323:Vcan protein, mouse; D016222:Fibroblast Growth Factor 2; D053675:Versicans; D020723:Glucuronic Acid; D003094:Collagen