| Abstract : |
The human urine is known to contain many components that potentially can serve as a source of biomarkers of health and disease. One of these important components are called exosomes (30-120 nm diameter) which are small biologically active nanoparticles that are secreted by the renal epithelial cells in the kidneys. Analysis of these exosomes by many groups showed that as these exosomes are fused to the membrane and released, many proteins tend to be released with the exosomes. A study by Pisitkun et. al, for example, has showed 295 different proteins found in urinary exosomes. Analyzing the kinds of proteins that are found in the urine can be used to classify and identify renal diseases. Urinary biomarkers can help in the process of identifying renal diseases and finding ways to treat these diseases. Understanding the importance of these exosomes is simply the first step and more progress can only be made when these exosomes are isolated and studies are conducted on them. Since research on exosomes has started, the major method used to extract these exosomes has been Ultracentrifugation (Pisitkun et. al). This project introduces and examines an additional protocol to isolate exosomes from urine to use for the identification of renal diseases. Normal and diabetic human pregnancy urine samples were taken and exosomes were isolated using the two methods and the exosomes were then imaged via electron microscopy. We concluded that the electron microscopy analysis of the exosomes shows that the exosomes from both the normal and diabetic from the two methods are comparable. The fact that there are no major differences between the exosomes from the two methods suggests that the method we propose can be used as a form of isolating exosomes quickly and efficiently. In the future this method can be used to isolate exosomes in a clinical laboratory and the exosomes can be used as a form of diagnosis and treatment. |