| RESEARCH PAPER
|Year : 2006 | Volume
| Issue : 1 | Page : 43-48
Inhibition of chondrogenic differentiation in chick limb-bud mesenchyme microcultures treated with cyclosporine
Soghra Bahmanpour1, Douglas F Paulsen2
1 Anatomy Department, Shiraz University of Medical Sciences, Shiraz, Iran
2 Departments of Anatomy & Neurobiology, Morehouse School of Medicine, Atlanta, GA, USA
Objectives: To explore the effects of cyclosporine (CsA) on skeletal development (chondrogenesis).
Materials and Methods: Mesenchymal cells obtained from stage-23 to stage-24 chick-embryo limb buds were grown in 96-well plates using chemically defined tissue-culture medium. Cultures were treated with CsA (0.01-5.0 µg/ml) and incubated (37ºC, 5% CO2) with daily medium changes for 4 days. After incubation of the cells in multiwell plate, cartilage differentiation (chondrogenesis) was assessed by selectively staining sulfated glycosaminoglycans (GAGs) in the cartilage matrix with Alcian blue, extracting the GAGs with 4 M guanidinium HCl, and spectrophotometric analysis of the extracts. Results: CsA treatment had concentration-dependent effects on chick limb-bud mesenchymal cell cultures. At 5 µg/ml, CsA caused cell loss, as judged microscopically by the paucity of cells remaining at the end of the culture period. CsA concentrations between 0.1 and 1 µg/ml caused a marked, dose-dependent decrease in chondrogenesis. At 0.01 µg/ml, CsA had no significant effect on chondrogenesis. At concentrations above 0.01 µg/ml, normalized data showed significant chondrogenic inhibition at 0.5 and 1.0 µg/ml CsA. Conclusions: The findings suggest a possible biological basis for CsA-associated effects on mesenchyme-derived tissues and provide a model system for further studies.
Anatomy Department, Shiraz University of Medical Sciences, Shiraz
Source of Support: None, Conflict of Interest: None
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