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| 1 | Methods for the extraction and RNA profiling of exosomes显示文摘AIM:To develop protocols for isolation of exosomes and characterization of their RNA content.METHODS:Exosomes were extracted from He La cell culture media and human blood serum using the Total exosome isolation(from cell culture media)reagent,and Total exosome isolation(from serum)reagent respectively.Identity and purity of the exosomes was confirmed by Nanosight?analysis,electron microscopy,and Western blots for CD63 marker.Exosomal RNA cargo was recovered with the Total exosome RNA and protein isolation kit.Finally,RNA was profiled using Bioanalyzer and quantitative reverse transcriptionpolymerase chain reaction(q RT-PCR)methodology.RESULTS:Here we describe a novel approach for robust and scalable isolation of exosomes from cell culture media and serum,with subsequent isolation and analysis of RNA residing within these vesicles.The isolation procedure is completed in a fraction of the time,compared to the current standard protocols utilizing ultracentrifugation,and allows to recover fully intact exosomes in higher yields.Exosomes were found tocontain a very diverse RNA cargo,primarily short sequences 20-200 nt(such as mi RNA and fragments of m RNA),however longer RNA species were detected as well,including full-length 18S and 28S r RNA.CONCLUSION:We have successfully developed a set of reagents and a workflow allowing fast and efficient extraction of exosomes,followed by isolation of RNA and its analysis by q RT-PCR and other techniques. | Emily Zeringer Mu Li Tim Barta Jeoffrey Schageman Ketil Winther Pedersen Axl Neurauter Susan Magdaleno Robert Setterquist Alexander V Vlassov | 2013 | World Journal of Methodology2013，3，1： | 5 |
| 2 | Structural and functional comparison of the RING domains of two p53 E3 ligases, Mdm2 and Pirh2显示文摘 | Shloush J Vlassov JE Engson I | 2011 | J Biol Chem2011，286，6： | 1 |
| 3 | Exosomes:current knowledge of their composition,biological functions,anddiagnostic and therapeutic potentials显示文摘 | Vlassov A V Magdaleno S Setterquist R | 2012 | Biochim Biophys Acta2012，1820，7： | 1 |
| 4 | Exosome , currrent knowledge of their composition, biological function, and diagnostic and therapeutic potentials 显示文摘 | Vlassov AV Magdaleno S Setterquist R | 2012 | Biochim Biophys Acta2012，1820，7： | 1 |
| 5 | Cationic lipid- DNA complexes-lipoplexes-for gene transfer and therapy 显示文摘 | ZHDANOV RI PODOBED OV VLASSOV VV | 2002 | Bioelectrochemistry2002，58，1： | 1 |
| 6 | Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials显示文摘 | Vlassov A V Magdaleno S Setterquist R | 2012 | Biochim Biopto s Acta2012，1820，7： | 1 |
| 7 | Circulating nucleic acids as a potential source for cancer biomarkers显示文摘 | Vlassov V V Laktionov P P Rykova E Y | 2010 | Curr Mol Med2010，10，2： | 1 |
| 8 | Canonic lipid-DNA complexes-lipoplexes for gene transfer and therapy显示文摘 | Zhdanov RI Podobed OV Vlassov VV | 2002 | Bioelectrochemistry2002，58，1： | 1 |
| 9 | Number of chronic conditions and professional longevity of aviators显示文摘 | Vlassov VV | 1997 | Aviat Space Environ Med1997，68，5： | 1 |
| 10 | Exosomes: cur- rent knowledge of their composition, biological functions, and di- agnostic and therapeutic potentials 显示文摘 | Vlassov AV Magdaleno S Setterquist R | 2012 | Biochim Biophys Acta2012，1820，7： | 1 |
| 11 | Number of chronic conditions and professional longevity of a aviators显示文摘 | Vlassov VV | 1997 | Aviat Space Environ Med1997，68，5： | 1 |
| 12 | Generalized extremal optimization for solving complex optimal design problems显示文摘 | Sousa F L Vlassov V Ramos F M | 2003 | Lecture Notes in Computer Science2003，2723，6： | 1 |
| 13 | Circulating nucleic acids as a potential source for cancer biomarkers显示文摘 | Vlassov VV Laktionov PP Rykova EY | 2010 | Curr Mol Med2010，10，2： | 1 |
| 14 | New approaches for cancer treatment: antitumor drugs based on gene-targeted nucleic acids显示文摘 | Patutina OA Mironova NL Vlassov VV | 2009 | Acta Naturae2009，1，2： | 1 |
| 15 | Totally endoscopic myocardial revascularization, an experimental study显示文摘 | Vlassov GP Tmvine N Deyneka KS | 1999 | Ann Thome Surg1999，68，4： | 1 |
| 16 | Heat pipc design through generalized extremal optimization 显示文摘 | Sousa F L Vlassov V Ramos F M | 2004 | Heat Transfer Engineering2004，25，7： | 1 |
| 17 | Generalized extremal optimization: An application in hcat pipe design显示文摘 | Sousa F L Vlassov V Ramos F M | 2004 | Applied Mathcmatical Modelling2004，28，10： | 1 |
| 18 | RNAi:a novel antisense technology and its therapeutic potential显示文摘 | Dallas A Vlassov A V | 2006 | Med Sci Monit2006，12，： | 1 |
| 19 | Exosomes:Current knowledge of their composition,biological functions,and diagnostic and therapeutic potentials显示文摘 | Vlassov A Magdaleno S Setterquist R | 2012 | Biochim Biophys Acta2012，1820，7： | 1 |
| 20 | Exosomes: Current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials显示文摘 | Alexander V. Vlassov Susan Magdaleno Robert Setterquist Rick Conrad | 2012 | BBA - General Subjects2012，，7： | 1 |
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