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Table 1 Procedure of exosome preparation [41]

From: Are exosomes the vehicle for protein aggregate propagation in neurodegenerative diseases?

1. Differential ultracentrifugation

Culture supernatant or fluid – at each of steps, the supernatant is used for following step.

⇒ 300 x g, 10 min (→ Pellet: cells)

⇒ 2000 x g, 10 min (→ Pellet: dead cells)

⇒ 10,000 x g, 30 min (→ Pellet: cell debris)

⇒ Final 100,000 x g, 70 min â†’ Pellet: exosomes + contaminating proteins

⇒ Wash in PBS 100,000 x g, 70 min → Pellet: correspond to exosomes

2. Rate-zonal centrifugation

- Load exosome preparation at the gradients (0.25 - 2 M sucrose) and centrifuge overnight (≥14 h) at 210,000 x g, 4 Â°C.

- With a micropipettor, collect eleven 1-mL fractions, from top to bottom.