Monoclonal antibody 11-fibrau: A useful marker to characterize chondrocyte differentiation stage

Department of Otorhinolaryngology, Erasmus University Medical Center Rotterdam, The Netherlands.
Biochemical and Biophysical Research Communications (Impact Factor: 2.3). 02/2001; 280(3):806-12. DOI: 10.1006/bbrc.2000.4168
Source: PubMed

ABSTRACT The aim of this study was to determine the feasibility of discriminating between differentiated and dedifferentiated chondrocytes by using the Mab 11-fibrau. Mab 11-fibrau did not bind to differentiated chondrocytes in cartilage of human knee joint, auricle, or nasal septum. During monolayer culture, when cells dedifferentiate, the number of 11-fibrau positive cells gradually increased and reached up to 100% after 4 passages. When differentiated chondrocytes were cultured in alginate, most (90--95%) of the cells remained 11-fibrau negative, in accordance with previous studies demonstrating that differentiated chondrocytes cultured in alginate keep their phenotype. Dedifferentiated (11-fibrau positive) cells were subjected to different redifferentiation regimes. As a well-known fact, cultures in alginate in medium where FCS was replaced by IGF1 and TGF beta 2 results in increased collagen type II formation, indicative for redifferentiation. However, the cells remained 11-fibrau positive, suggesting they are not (yet) fully redifferentiated. On the other hand, when dedifferentiated cells (after 4 passages in monolayer culture) were seeded in a biomaterial and implanted subcutaneously in a nude mouse, the newly formed cartilage matrix contained collagen type II and the 11-fibrau staining on the cells had disappeared. Our results indicate that 11-fibrau may be a reliable and sensitive marker of chondrocyte phenotype.

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    • "Cells and histological sections were incubated with either mouse monoclonal antibody against 11-fibrau (Clone D7-FIB; diluted 1:400; Imgen, Netherlands), a marker for fibroblasts [19], or monoclonal antibody against α-SMA (Clone 1A4; diluted 1:1000; Sigma, St.Louis, Missouri, USA), a marker for smooth muscle cells and pericytes [20], for two hours. Cells were rinsed in 1×PBS and IHC detection was performed using Link-Label (Biotin-based) Multilink® IHC Detection Kit (Biogenex, San Ramon, CA). "
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