caa a22 4500 606155058 CHVBK 20210128100601.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s10561-014-9472-7 doi (NATIONALLICENCE)springer-10.1007/s10561-014-9472-7 Alternative protocols to induce chondrogenic differentiation: transforming growth factor-β superfamily [Elektronische Daten] [Claudia Cicione, Emma Muiños-López, Tamara Hermida-Gómez, Isaac Fuentes-Boquete, Silvia Díaz-Prado, Francisco Blanco] 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. Springer Science+Business Media Dordrecht, 2014 Cell therapy nationallicence Autologous chondrocyte implantation nationallicence Focal lesions nationallicence Hyaline articular cartilage nationallicence AA : Ascorbic acid nationallicence AGG : Aggrecan nationallicence ALP : Alkaline phosphatase nationallicence APM1 : Adiponectin nationallicence BM-MSCs : Bone marrow-mesenchymal stem cells nationallicence BMP-2 : Bone morphogenetic protein-2 nationallicence BMP-7 : Bone morphogenetic protein-7 nationallicence cDNA : Complementary deoxyribonucleic acid nationallicence COL10A1 : Collagen type XA1 nationallicence COL1A1 : Collagen type IA1 nationallicence COL2A1 : Collagen type IIA1 nationallicence DMEM : Dulbecco's modified Eagle's medium nationallicence FABP4 : Fatty acid-binding protein 4 nationallicence FBS : Fetal bovine serum nationallicence HE : Hematoxylin-eosin nationallicence IGF-1 : Insulin-like growth factor-1 nationallicence KO : Knockout serum nationallicence LPL : Lipoprotein lipase nationallicence MMP13 : Matrix metalloproteinase 13 nationallicence MSCs : Mesenchymal stem cells nationallicence MT : Masson's trichrome nationallicence MTG : Monotioglycerol nationallicence OP : Osteoprotegerin nationallicence OP-1 : Osteogenic protein 1 nationallicence P/S : Penicillin and streptomycin nationallicence PCR : Polymerase chain reaction nationallicence qPCR : Real-Time PCR nationallicence REL : Relative expression levels nationallicence rHuBMP-2 : Recombinant human bone morphogenetic protein 2 nationallicence RNA : Ribonucleic acid nationallicence SaO : Safranin O nationallicence SOX9 : Sex determining region Y-box 9 nationallicence TB : Toluidine blue nationallicence TBP : TATA box binding protein nationallicence TGFβ : Transforming growth factor beta nationallicence TGFβ-3 : Transforming growth factor beta-3 nationallicence Cicione Claudia División de Reumatología, INIBIC-Hospital Universitario A Coruña, C/As Xubias S/N, 15.006, A Coruña, Spain aut Muiños-López Emma División de Reumatología, INIBIC-Hospital Universitario A Coruña, C/As Xubias S/N, 15.006, A Coruña, Spain aut Hermida-Gómez Tamara División de Reumatología, INIBIC-Hospital Universitario A Coruña, C/As Xubias S/N, 15.006, A Coruña, Spain aut Fuentes-Boquete Isaac Servicio de Reumatología, Grupo de Bioingeniería Tisular y Terapia Celular (CBTTC-CHUAC), CIBER-BBN/ISCIII, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), SERGAS, Universidade da Coruña (UDC), As Xubias, 15.006, A Coruña, Spain aut Díaz-Prado Silvia Servicio de Reumatología, Grupo de Bioingeniería Tisular y Terapia Celular (CBTTC-CHUAC), CIBER-BBN/ISCIII, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), SERGAS, Universidade da Coruña (UDC), As Xubias, 15.006, A Coruña, Spain aut Blanco Francisco División de Reumatología, INIBIC-Hospital Universitario A Coruña, C/As Xubias S/N, 15.006, A Coruña, Spain aut Cell and Tissue Banking Springer Netherlands 16/2(2015-06-01), 195-207 1389-9333 16:2<195 2015 16 10561 https://doi.org/10.1007/s10561-014-9472-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10561-014-9472-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Cicione Claudia División de Reumatología, INIBIC-Hospital Universitario A Coruña, C/As Xubias S/N, 15.006, A Coruña, Spain aut NATIONALLICENCE 700 1- Muiños-López Emma División de Reumatología, INIBIC-Hospital Universitario A Coruña, C/As Xubias S/N, 15.006, A Coruña, Spain aut NATIONALLICENCE 700 1- Hermida-Gómez Tamara División de Reumatología, INIBIC-Hospital Universitario A Coruña, C/As Xubias S/N, 15.006, A Coruña, Spain aut NATIONALLICENCE 700 1- Fuentes-Boquete Isaac Servicio de Reumatología, Grupo de Bioingeniería Tisular y Terapia Celular (CBTTC-CHUAC), CIBER-BBN/ISCIII, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), SERGAS, Universidade da Coruña (UDC), As Xubias, 15.006, A Coruña, Spain aut NATIONALLICENCE 700 1- Díaz-Prado Silvia Servicio de Reumatología, Grupo de Bioingeniería Tisular y Terapia Celular (CBTTC-CHUAC), CIBER-BBN/ISCIII, Instituto de Investigación Biomédica de A Coruña (INIBIC), Complexo Hospitalario Universitario de A Coruña (CHUAC), SERGAS, Universidade da Coruña (UDC), As Xubias, 15.006, A Coruña, Spain aut NATIONALLICENCE 700 1- Blanco Francisco División de Reumatología, INIBIC-Hospital Universitario A Coruña, C/As Xubias S/N, 15.006, A Coruña, Spain aut NATIONALLICENCE 773 0- Cell and Tissue Banking Springer Netherlands 16/2(2015-06-01), 195-207 1389-9333 16:2<195 2015 16 10561