tailieunhanh - báo cáo hóa học:" Body fluid derived exosomes as a novel template for clinical diagnostics"

Tuyển tập các báo cáo nghiên cứu về hóa học được đăng trên tạp chí sinh học đề tài : Body fluid derived exosomes as a novel template for clinical diagnostics | Keller et al. Journal of Translational Medicine 2011 9 86 http content 9 1 86 JOURNAL OF TRANSLATIONAL MEDICINE RESEARCH Open Access Body fluid derived exosomes as a novel template for clinical diagnostics 2 2 2 1 2 Sascha Keller Johannes Ridinger Anne-Kathleen Rupp Johannes WG Janssen and Peter Altevogt Abstract Background Exosomes are small membrane vesicles with a size of 40-100 nm that are released by different cell types from a late endosomal cellular compartment. They can be found in various body fluids including plasma malignant ascites urine amniotic fluid and saliva. Exosomes contain proteins miRNAs and mRNAs exosome shuttle RNA esRNA that could serve as novel platform for diagnosis. Method We isolated exosomes from amniotic fluid saliva and urine by differential centrifugation on sucrose gradients. Marker proteins were identified by Western blot and FACS analysis after adsorption of exosomes to latex beads. We extracted esRNA from exosomes carried out RT-PCR and analyzed amplified products by restriction length polymorphism. Results Exosomes were positive for the marker proteins CD24 CD9 Annexin-1 and Hsp70 and displayed the correct buoyant density and orientation of antigens. In sucrose gradients the exosomal fractions contained esRNA that could be isolated with sufficient quantity for further analysis. EsRNAs were protected in exosomes from enzymatic degradation. Amniotic fluid esRNA served as template for the typing of the CD24 single nucleotide polymorphism rs52812045 . It also allowed sex determination of the fetus based on the detection of the male specific ZFY gene product. Conclusions Our data demonstrate that exosomes from body fluids carry esRNAs which can be analyzed and offers access to the transcriptome of the host organism. The exosomal lipid bilayer protects the genetic information from degradation. As the isolation of exosomes is a minimally invasive procedure this technique opens new possibilities for .