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Housekeeping gene stability influences the quantification of osteogenic markers during stem cell differentiation to the osteogenic lineage / Carlos Augusto Sarassa Velásquez  

Público ISBD Título : Housekeeping gene stability influences the quantification of osteogenic markers during stem cell differentiation to the osteogenic lineage Tipo de documento : documento electrónico Autores : Carlos Augusto Sarassa Velásquez, Fecha de publicación : 2010 Títulos uniformes : Cytotechnology Idioma : Inglés (eng) Palabras clave : Mesenchymal stem cells  Osteogenic differentiation  Normalization  Gene expression  Quantitative RT-PCR Resumen : Real-time reverse transcription PCR (RT-qPCR) relies on a housekeeping or normalizer gene whose expression remains constant throughout the experiment. RT-qPCR is commonly used for characterization of human bone marrow mesenchymal stem cells (hBMSCs). However, to the best of our knowledge, there are no studies validating the expression stability of the genes used as normalizers during hBMSCs differentiation. This work aimed to study the stability of the housekeeping genes β-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ribosomal protein L13A (RPL13A) during the osteogenic differentiation of hBMSCs. Their stability was evaluated via RT-qPCR in 14 and 20 day differentiation assays to the osteogenic lineage. Different normalization strategies were evaluated to quantify the osteogenic markers collagen type I, bone sialoprotein and osteonectin. Cell differentiation was confirmed via alizarin red staining. The results demonstrated up-regulation of β-actin with maximum fold changes (MFC) of 4.38. GAPDH and RPL13A were not regulated by osteogenic media after 14 days and presented average fold changes lower than 2 in 20 day cultures. RPL13A (MFC Mención de responsabilidad : Felipe Garcia Quiroz, Olga M Posada, Daniel Gallego-Perez, Natalia Higuita-Castro, Carlos Sarassa, Derek J Hansford, Piedad Agudelo-Florez, Luis E López Referencia : Cytotechnology. 2010 Apr;62(2):109-20. DOI (Digital Object Identifier) : 10.1007/s10616-010-9265-1 PMID : 20396946 En línea : http://link.springer.com/10.1007/s10616-010-9265-1 Enlace permanente : https://hospitalpablotobon.cloudbiteca.com/pmb/opac_css/index.php?lvl=notice_display&id=4468 Housekeeping gene stability influences the quantification of osteogenic markers during stem cell differentiation to the osteogenic lineage [documento electrónico] / Carlos Augusto Sarassa Velásquez,  . - 2010. Obra : Cytotechnology Idioma : Inglés (eng) Palabras clave : Mesenchymal stem cells  Osteogenic differentiation  Normalization  Gene expression  Quantitative RT-PCR Resumen : Real-time reverse transcription PCR (RT-qPCR) relies on a housekeeping or normalizer gene whose expression remains constant throughout the experiment. RT-qPCR is commonly used for characterization of human bone marrow mesenchymal stem cells (hBMSCs). However, to the best of our knowledge, there are no studies validating the expression stability of the genes used as normalizers during hBMSCs differentiation. This work aimed to study the stability of the housekeeping genes β-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ribosomal protein L13A (RPL13A) during the osteogenic differentiation of hBMSCs. Their stability was evaluated via RT-qPCR in 14 and 20 day differentiation assays to the osteogenic lineage. Different normalization strategies were evaluated to quantify the osteogenic markers collagen type I, bone sialoprotein and osteonectin. Cell differentiation was confirmed via alizarin red staining. The results demonstrated up-regulation of β-actin with maximum fold changes (MFC) of 4.38. GAPDH and RPL13A were not regulated by osteogenic media after 14 days and presented average fold changes lower than 2 in 20 day cultures. RPL13A (MFC Mención de responsabilidad : Felipe Garcia Quiroz, Olga M Posada, Daniel Gallego-Perez, Natalia Higuita-Castro, Carlos Sarassa, Derek J Hansford, Piedad Agudelo-Florez, Luis E López Referencia : Cytotechnology. 2010 Apr;62(2):109-20. DOI (Digital Object Identifier) : 10.1007/s10616-010-9265-1 PMID : 20396946 En línea : http://link.springer.com/10.1007/s10616-010-9265-1 Enlace permanente : https://hospitalpablotobon.cloudbiteca.com/pmb/opac_css/index.php?lvl=notice_display&id=4468

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