BACKGROUND AIMS: Stem cells derived from human adipose tissue (ASC) have the capacity for renewal, are easily obtained and have plasticity properties that allow them to differentiate into several cell types, including osteoblast cells. With the aim of understanding the issue of the osteogenic process and finding reliable biomarkers in cells undergoing the osteogeneic differentiation process, this work took advantage of a proteomic approach to identify proteins involved in osteogenesis. METHODS: For this purpose, ASC were analyzed under three conditions: S0, in the absence of stimulation; S1, with 2 weeks of osteogenic medium stimulation; and S2, with 4 weeks of osteogenic medium stimulation. The identification of ASC was carried out by flow cytometry using antibodies specific to known undifferentiated stem cell-surface markers. Cell viability, enzymatic activity, mineral deposition, collagen structure and production and gene analyzes were evaluated for each condition. RESULTS: Phenotypic modifications were observed during the in vitro osteogenic differentiation process by two-dimensional (2-D) differential image gel electrophoresis (DIGE). The proteins were identified by mass espectrometry in tandem (MS/MS) analyzes using Matrix-assisted laser desorption/ionization with TOF/TOF is a tandem mass spectrometry method where two time-of-flight mass spectrometers are used consecutively (MALDI-TOF/TOF). A total of 51 differentially expressed proteins was identified when comparing the three observed conditions. Sixteen different spots were identified in the S0 stage compared with S2, while 28 different spots were found in S2 compared with S0. S1 expressed seven different spots compared with S0 and S2. CONCLUSIONS: These findings suggest the involvement of several proteins directly related to the osteogenic pathway, which can be used to improve understanding of the osteogenic process.
BACKGROUND AIMS: Stem cells derived from human adipose tissue (ASC) have the capacity for renewal, are easily obtained and have plasticity properties that allow them to differentiate into several cell types, including osteoblast cells. With the aim of understanding the issue of the osteogenic process and finding reliable biomarkers in cells undergoing the osteogeneic differentiation process, this work took advantage of a proteomic approach to identify proteins involved in osteogenesis. METHODS: For this purpose, ASC were analyzed under three conditions: S0, in the absence of stimulation; S1, with 2 weeks of osteogenic medium stimulation; and S2, with 4 weeks of osteogenic medium stimulation. The identification of ASC was carried out by flow cytometry using antibodies specific to known undifferentiated stem cell-surface markers. Cell viability, enzymatic activity, mineral deposition, collagen structure and production and gene analyzes were evaluated for each condition. RESULTS: Phenotypic modifications were observed during the in vitro osteogenic differentiation process by two-dimensional (2-D) differential image gel electrophoresis (DIGE). The proteins were identified by mass espectrometry in tandem (MS/MS) analyzes using Matrix-assisted laser desorption/ionization with TOF/TOF is a tandem mass spectrometry method where two time-of-flight mass spectrometers are used consecutively (MALDI-TOF/TOF). A total of 51 differentially expressed proteins was identified when comparing the three observed conditions. Sixteen different spots were identified in the S0 stage compared with S2, while 28 different spots were found in S2 compared with S0. S1 expressed seven different spots compared with S0 and S2. CONCLUSIONS: These findings suggest the involvement of several proteins directly related to the osteogenic pathway, which can be used to improve understanding of the osteogenic process.
Authors: Alexandre De la Fuente; Jesús Mateos; Iván Lesende-Rodríguez; Valentina Calamia; Isaac Fuentes-Boquete; Francisco J de Toro; Maria C Arufe; Francisco J Blanco Journal: Mol Cell Proteomics Date: 2011-10-17 Impact factor: 5.911
Authors: Mika Pietilä; Sami Palomäki; Siri Lehtonen; Ilja Ritamo; Leena Valmu; Johanna Nystedt; Saara Laitinen; Hannnu-Ville Leskelä; Raija Sormunen; Juha Pesälä; Katrina Nordström; Ari Vepsäläinen; Petri Lehenkari Journal: Stem Cells Dev Date: 2011-07-20 Impact factor: 3.272
Authors: Laura Kyllönen; Suvi Haimi; Bettina Mannerström; Heini Huhtala; Kristiina M Rajala; Heli Skottman; George K Sándor; Susanna Miettinen Journal: Stem Cell Res Ther Date: 2013-02-15 Impact factor: 6.832
Authors: Mehran Dadras; Caroline May; Johannes Maximilian Wagner; Christoph Wallner; Mustafa Becerikli; Stephanie Dittfeld; Bettina Serschnitzki; Lukas Schilde; Annika Guntermann; Christina Sengstock; Manfred Köller; Dominik Seybold; Jan Geßmann; Thomas Armin Schildhauer; Marcus Lehnhardt; Katrin Marcus; Björn Behr Journal: J Cell Mol Med Date: 2020-09-03 Impact factor: 5.310