caa a22 4500 606236570 CHVBK 20210128101248.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s11010-014-2276-z doi (NATIONALLICENCE)springer-10.1007/s11010-014-2276-z Endothelial progenitor cells proliferated via MEK-dependent p42 MAPK signaling pathway [Elektronische Daten] [Ferry Sandra, Yudi Oktaviono, Mohammad Widodo, Yanni Dirgantara, Angliana Chouw, Djanggan Sargowo] Endothelial progenitor cells (EPCs) clinical applications have been well reported. However, due to low number of EPCs that could be isolated, EPCs expansion study became one of the main focuses. Some optimized mediums to culture EPCs were currently available. However, the proliferation signaling pathway is not clearly disclosed yet. Peripheral blood was collected from eight healthy subjects, followed by mononuclear cells (MNCs) isolation. MNCs were then prepared and cultured for 2days. After that, non-adherent cells were harvested and further cultured for 3days. Resulted colony-forming unit (CFU)-Hill colonies were documented and enumerated under an inverted light microscope. To detect membrane markers, immunofluorescence was performed to detect CD34, VEGFR-2, and CD133. Cell documentation was conducted under a fluorescence microscope. To check cell proliferation, XTT Cell Proliferation Assay Kit was used according to kit insert. To detect possible activation of p44/42 MAPK, western blot was performed to detect p44/42 MAPK and phosphorylated p44/42 MAPK. All visualized bands were captured and quantified. Our results showed that EPCs markers (CD34, CD133 and VEGFR-2) were detected in 3days culture. From XTT cell proliferation assay and CFU enumeration results, we found that EPCs proliferated significantly (p=0.012) with addition of supplement. Phosphorylated-p42 MAPK expression of EPCs treated with supplement was significantly higher than the one of EPCs without treatment. Significant inhibition of p42 MAPK phosphorylation by U0126 was observed (p=0.012). By pretreatment of U0126, number of viable cells and CFUs treated with supplement was significantly decreased (p=0.012). Our results showed that MEK-dependent p42 MAPK pathway might play an important role in EPCs proliferation. Springer Science+Business Media New York, 2014 Endothelial progenitor cell nationallicence EPC nationallicence p42 nationallicence Erk2 nationallicence MAPK nationallicence Proliferation nationallicence Sandra Ferry Prodia Clinical Laboratory, Prodia Tower, Jl. Kramat Raya No. 150, Jakarta, Indonesia aut Oktaviono Yudi Department of Cardiology and Vascular Medicine, Faculty of Medicine, Dr. Soetomo Hospital, University of Airlangga, Jl. May. Jend. Prof. Dr. Moestopo No. 6-8, Surabaya, Indonesia aut Widodo Mohammad Postgraduate Program in Biomedical Sciences, Faculty of Medicine, Brawijaya University, Jl. Veteran, Malang, Indonesia aut Dirgantara Yanni PT. Prodia Stemcell Indonesia, Jl. Kramat 7 No. 11, Jakarta, Indonesia aut Chouw Angliana PT. Prodia Stemcell Indonesia, Jl. Kramat 7 No. 11, Jakarta, Indonesia aut Sargowo Djanggan Postgraduate Program in Biomedical Sciences, Faculty of Medicine, Brawijaya University, Jl. Veteran, Malang, Indonesia aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 400/1-2(2015-02-01), 201-206 0300-8177 400:1-2<201 2015 400 11010 https://doi.org/10.1007/s11010-014-2276-z 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/s11010-014-2276-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sandra Ferry Prodia Clinical Laboratory, Prodia Tower, Jl. Kramat Raya No. 150, Jakarta, Indonesia aut NATIONALLICENCE 700 1- Oktaviono Yudi Department of Cardiology and Vascular Medicine, Faculty of Medicine, Dr. Soetomo Hospital, University of Airlangga, Jl. May. Jend. Prof. Dr. Moestopo No. 6-8, Surabaya, Indonesia aut NATIONALLICENCE 700 1- Widodo Mohammad Postgraduate Program in Biomedical Sciences, Faculty of Medicine, Brawijaya University, Jl. Veteran, Malang, Indonesia aut NATIONALLICENCE 700 1- Dirgantara Yanni PT. Prodia Stemcell Indonesia, Jl. Kramat 7 No. 11, Jakarta, Indonesia aut NATIONALLICENCE 700 1- Chouw Angliana PT. Prodia Stemcell Indonesia, Jl. Kramat 7 No. 11, Jakarta, Indonesia aut NATIONALLICENCE 700 1- Sargowo Djanggan Postgraduate Program in Biomedical Sciences, Faculty of Medicine, Brawijaya University, Jl. Veteran, Malang, Indonesia aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 400/1-2(2015-02-01), 201-206 0300-8177 400:1-2<201 2015 400 11010