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Urine exosome preparation

MS Masaki Sakai
KY Keiko Yamamoto
HM Hiroaki Mizumura
TM Tomoki Matsumoto
YT Yasuko Tanaka
YN Yumi Noda
KI Kenichi Ishibashi
TY Tadashi Yamamoto
SS Sei Sasaki
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Exosomes were prepared from urine samples by an ultracentrifugation method [12]. In brief, urine samples were centrifuged at 3,000 × g for 15 min at 25 °C to remove sediments, debris and cells. Then, the supernatants were further centrifuged at 17,000 × g for 15 min at 25 °C to remove larger vesicles. The supernatants were finally ultracentrifuged at 160,000 or 200,000 × g for 1 h at 25 °C to collect exosomes. The exosomes were suspended in phosphate buffered saline for further analysis. In human, urine samples were collected from the first urine in the morning from healthy volunteers who had given written informed consent (Ethical Committee of Meiji Pharmaceutical University; approved number 2609). Mouse urine samples were obtained from the stored urine collected by metabolic cages.

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