title: Microfluidic isolation and transcriptome analysis of serum microvesicles.
abstract: 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.
authors.affiliation: BioMEMS Resource Center, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA.
authors.forename: Chihchen
authors.initials: C
authors.lastname: Chen
authors.forename: Johan
authors.initials: J
authors.lastname: Skog
authors.forename: Chia-Hsien
authors.initials: CH
authors.lastname: Hsu
authors.forename: Ryan T
authors.initials: RT
authors.lastname: Lessard
authors.forename: Leonora
authors.initials: L
authors.lastname: Balaj
authors.forename: Thomas
authors.initials: T
authors.lastname: Wurdinger
authors.forename: Bob S
authors.initials: BS
authors.lastname: Carter
authors.forename: Xandra O
authors.initials: XO
authors.lastname: Breakefield
authors.forename: Mehmet
authors.initials: M
authors.lastname: Toner
authors.forename: Daniel
authors.initials: D
authors.lastname: Irimia
mesh_terms: 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
chemical_list: D017077:Culture Media, Conditioned; D012313:RNA