Microfluidic isolation and transcriptome analysis of serum microvesicles

被引:448
作者
Chen, Chihchen [1 ]
Skog, Johan [2 ,3 ]
Hsu, Chia-Hsien [1 ]
Lessard, Ryan T. [2 ,3 ]
Balaj, Leonora [2 ,3 ]
Wurdinger, Thomas [2 ,3 ,4 ]
Carter, Bob S. [5 ]
Breakefield, Xandra O. [2 ,3 ]
Toner, Mehmet [1 ]
Irimia, Daniel [1 ]
机构
[1] Harvard Univ, Sch Med, Shriners Hosp Children,Massachusets Gen Hosp, BioMEMS Resource Ctr,Ctr Engn Med & Surg Serv, Boston, MA 02114 USA
[2] Harvard Univ, Sch Med, Massachusetts Gen Hosp, Neurosci Program,Dept Neurol, Boston, MA 02114 USA
[3] Harvard Univ, Sch Med, Massachusetts Gen Hosp, Neurosci Program,Dept Radiol, Boston, MA 02114 USA
[4] Vrije Univ Amsterdam, Med Ctr, Dept Neurosurg, Neurooncol Res Grp, NL-1007 MB Amsterdam, Netherlands
[5] Harvard Univ, Sch Med, Massachusetts Gen Hosp, Dept Neurosurg, Boston, MA 02114 USA
关键词
TUMOR-DERIVED EXOSOMES; POLYMERASE CHAIN-REACTION; DENDRITIC CELLS; DIAGNOSTIC BIOMARKERS; MAJOR INHIBITOR; PLASMA-MEMBRANE; CANCER; BLOOD; IDENTIFICATION; AMPLIFICATION;
D O I
10.1039/b916199f
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Microvesicles (exosomes) shed from both normal and cancerous cells may serve as means of intercellular communication. These microvesicles carry proteins, lipids and nucleic acids derived from the host cell. Their isolation and analysis from blood samples have the potential to provide information about state and progression of malignancy and should prove of great clinical importance as biomarkers for a variety of disease states. However, current protocols for isolation of microvesicles from blood require high-speed centrifugation and filtration, which are cumbersome and time consuming. In order to take full advantage of the potential of microvesicles as biomarkers for clinical applications, faster and simpler methods of isolation will be needed. In this paper, we present an easy and rapid microfluidic immunoaffinity method to isolate microvesicles from small volumes of both serum from blood samples and conditioned medium from cells in culture. RNA of high quality can be extracted from these microvesicles providing a source of information about the genetic status of tumors to serve as biomarkers for diagnosis and prognosis of cancer.
引用
收藏
页码:505 / 511
页数:7
相关论文
共 47 条
[1]  
Abu Al-Soud W, 2000, J CLIN MICROBIOL, V38, P345
[2]   Neutrophil migration assay from a drop of blood [J].
Agrawal, Nitin ;
Toner, Mehmet ;
Irimia, Daniel .
LAB ON A CHIP, 2008, 8 (12) :2054-2061
[3]  
AKANE A, 1994, J FORENSIC SCI, V39, P362
[4]   Intercellular transfer of the oncogenic receptor EGFrvIII by microvesicles derived from tumour cells [J].
Al-Nedawi, Khalid ;
Meehan, Brian ;
Micallef, Johann ;
Lhotak, Vladimir ;
May, Linda ;
Guha, Abhijit ;
Rak, Janusz .
NATURE CELL BIOLOGY, 2008, 10 (05) :619-U24
[5]   Exosomes as potent cell-free peptide-based vaccine.: I.: Dendritic cell-derived exosomes transfer functional MHC class I/peptide complexes to dendritic cells [J].
André, F ;
Chaput, N ;
Schartz, NEC ;
Flament, C ;
Aubert, N ;
Bernard, J ;
Lemonnier, F ;
Raposo, G ;
Escudier, B ;
Hsu, DH ;
Tursz, T ;
Amigorena, S ;
Angevin, E ;
Zitvogel, L .
JOURNAL OF IMMUNOLOGY, 2004, 172 (04) :2126-2136
[6]   Malignant effusions and immunogenic tumour-derived exosomes [J].
Andre, F ;
Schartz, NEC ;
Movassagh, M ;
Flament, C ;
Pautier, P ;
Morice, P ;
Pomel, C ;
Lhomme, C ;
Escudier, B ;
Le Chevalier, T ;
Tursz, T ;
Amigorena, S ;
Raposo, G ;
Angevin, E ;
Zitvogel, L .
LANCET, 2002, 360 (9329) :295-305
[7]   Exosomes and HIV Gag bud from endosome-like domains of the T cell plasma membrane [J].
Booth, AM ;
Fang, Y ;
Fallon, JK ;
Yang, JM ;
Hildreth, JEK ;
Gould, SJ .
JOURNAL OF CELL BIOLOGY, 2006, 172 (06) :923-935
[8]   Discrimination between exosomes and HIV-1:: Purification of both vesicles from cell-free supernatants [J].
Cantin, Rejean ;
Diou, Juliette ;
Belanger, Dave ;
Tremblay, Alexandre M. ;
Gilbert, Caroline .
JOURNAL OF IMMUNOLOGICAL METHODS, 2008, 338 (1-2) :21-30
[9]   A microfluidic device for practical label-free CD4+T cell counting of HIV-infected subjects [J].
Cheng, Xuanhong ;
Irimia, Daniel ;
Dixon, Meredith ;
Sekine, Kazuhiko ;
Demirci, Utkan ;
Zamir, Lee ;
Tompkins, Ronald G. ;
Rodriguez, William ;
Toner, Mehmet .
LAB ON A CHIP, 2007, 7 (02) :170-178
[10]   Continuous hydrophoretic separation and sizing of microparticles using slanted obstacles in a microchannel [J].
Choi, Sungyoung ;
Park, Je-Kyun .
LAB ON A CHIP, 2007, 7 (07) :890-897