Alternative protocols to induce chondrogenic differentiation: transforming growth factor-β superfamily


Mesenchymal stem cells (MSCs) are an accepted candidate for cell-based therapy of multiple diseases. The interest in MSCs and their possible application in cell therapy have resulted in a better understanding of the basic biology of these cells. Recently, like aggregation and transforming growth factor beta (TGFβ) delivery, hypoxia has been indicated as crucial for complete chondrogenesis. The aim of this study was to test different culture conditions for directing stem cell differentiation into the chondrogenic lineage in vitro by testing different TGFβ superfamily members into the culture media under normoxic conditions. All chondrogenic culture conditions used allowed the differentiation of bone marrow-MSCs (BM-MSCs) into chondrogenic lineage. Chondrogenic induction capacity depended on the growth factor added to the culture media. In particular, the chondrogenic culture condition that better induced chondrogenesis was the medium that included the combination of three growth factors: bone morphogenetic protein-2 (BMP-2), BMP-7 and TGFβ-3. In this culture media, differentiated cells showed the highest levels expression of two markers of chondrogenesis, SOX9 and COL2A1, compared to the control points (p < 0.05, two-tailed t test). In our experimental conditions, the combination of BMP-2, BMP-7 and TGFβ-3 was the most effective in promoting chondrogenesis of BM-MSCs. These results underline the importance of determining in each experimental design the best protocol for in vitro directing stem cell differentiation into the chondrogenic lineage.

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Ascorbic acid




Alkaline phosphatase




Bone marrow-mesenchymal stem cells


Bone morphogenetic protein-2


Bone morphogenetic protein-7


Complementary deoxyribonucleic acid


Collagen type XA1


Collagen type IA1


Collagen type IIA1


Dulbecco’s modified Eagle’s medium


Fatty acid-binding protein 4


Fetal bovine serum




Insulin-like growth factor-1


Knockout serum


Lipoprotein lipase


Matrix metalloproteinase 13


Mesenchymal stem cells


Masson’s trichrome






Osteogenic protein 1


Penicillin and streptomycin


Polymerase chain reaction


Real-Time PCR


Relative expression levels


Recombinant human bone morphogenetic protein 2


Ribonucleic acid


Safranin O


Sex determining region Y-box 9


Toluidine blue


TATA box binding protein


Transforming growth factor beta


Transforming growth factor beta-3


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This study was supported by grants: Servizo Galego de Saúde, Xunta de Galicia (PS07/84), Cátedra Bioiberica de la Universidade da Coruña and Instituto de Salud Carlos III CIBER BBN; Ministerio Ciencia e Innovacion PLE2009-0144; Fondo Investigacion Sanitaria-PI 08/2028 with participation of funds from FEDER (European Community), Tamara Hermida-Gómez is the beneficiary of a contract from Fondo de Investigación Sanitaria (2008), Spain. We would like to thank P.Filgueira and M.J.Sánchez for technical assistance.

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The authors declare that no competing financial interest exists.

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Correspondence to Silvia Díaz-Prado or Francisco J. Blanco.

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Cicione, C., Muiños-López, E., Hermida-Gómez, T. et al. Alternative protocols to induce chondrogenic differentiation: transforming growth factor-β superfamily. Cell Tissue Bank 16, 195–207 (2015).

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  • Cell therapy
  • Autologous chondrocyte implantation
  • Focal lesions
  • Hyaline articular cartilage