Regulation of Chondrogenesis by Transforming Growth Factor-β3 and Insulin-like Growth Factor-1 from Human Mesenchymal Umbilical Cord Blood Cells

Objective. Mature articular cartilage is vulnerable to injuries and disease processes that cause irreversible tissue damage because of its limited capacity for self-repair. Umbilical cord blood is a source of mesenchymal stem cells, which can give rise to cells of different lineages, including cartilage, bone, and fat. Cellular condensation is a required step in the initiation of mesenchymal chondrogenesis. We attempted to differentiate cells from umbilical cord blood into chondrocytes with insulin-like growth factor 1 (IGF-1) and transforming growth factor-beta 3 (TGF-beta 3). Methods. Cells were grown in high density micromass and monolayer culture systems and then evaluated for expression of type 11 collagen, aggrecan, and Sox9. Umbilical cord blood from 130 patients was harvested. Results. Expression of type II collagen, aggrecan, and Sox9 was detected after 14 days in TGF-beta 3- and IGF-1-stimulated cells in both types of culture (monolayer and micromass). On Day 21 in the micromass culture, expression levels were greater than they were at 14 days for all genes. TGF-beta 3 was found to be more efficient at promoting chondrogenesis than IGF-1. By western blot, we also found that after 3 weeks, the expression of type II collagen was greater in micromass culture with TGF-beta 3. Conclusion. TGF-beta 3 used in micromass culture is the best growth factor for promoting the proliferation and differentiation of mesenchymal cells from umbilical cord blood during chondrogenesis. This approach may provide an alternative to autologous grafting. (First Release May 15 2010; J Rheumatol 2010;37:1519-26; doi: 10.3899/jrheum.091169)