Microfluidic isolation and transcriptome analysis of serum microvesicles.
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.
Chen
Chihchen
C
BioMEMS Resource Center, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA.
Skog
Johan
J
Hsu
Chia-Hsien
CH
Lessard
Ryan T
RT
Balaj
Leonora
L
Wurdinger
Thomas
T
Carter
Bob S
BS
Breakefield
Xandra O
XO
Toner
Mehmet
M
Irimia
Daniel
D
D000915:Antibody Affinity; D045744:Cell Line, Tumor; D002474:Cell-Free System / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000378:metabolism; D055354:Exosomes / Q000378:metabolism*; D020869:Gene Expression Profiling; D006801:Humans; D046210:Microfluidic Analytical Techniques; D009369:Neoplasms / Q000175:diagnosis; D019095:Point-of-Care Systems; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D044967:Serum / Q000166:cytology*; D013499:Surface Properties; D013997:Time Factors
D017077:Culture Media, Conditioned; D012313:RNA