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. Author manuscript; available in PMC: 2015 Jun 2.
Published in final edited form as: Kidney Int. 2010 Apr 28;78(2):191–199. doi: 10.1038/ki.2010.106

Figure 6. Isolation of microvesicles using filtration concentrators reveals that this may be a rapid technique to isolate intact microvesicles for nucleic acid extraction.

Figure 6

(a) A sample of 75 ml human urine was subjected to the initial processing steps of 300 g, 17,000 g centrifugation followed by 0.8 µm filtration and was then processed using ultracentrifugation (blue) or using 100 kDa MWCO filters (red). The results revealed that a similar profile was obtained using both extraction methods with minimal degradation of the RNA revealing that filtration concentrators may be a fast and reliable way to isolate urinary microvesicles. Comparison of the ‘normal’ 300 g, 17,000 g spin and 0.8 µm filtration steps (red) with just a 0.8 µm filtration step (blue) followed by (b) ultracentrifugation or (c) filtration concentrators revealed that the 300 g and 17,000 g spins were not crucial for the removal of cell debris and whole cells as no change in profile was observed, further simplifying the isolation technique and the time taken to isolate exosomes. All samples underwent extra-exosomal RNase and DNase digestion before microvesicular lysis.