A method for isolating exosomes from stem cells using single-use bioreactors and high-speed/ultracentrifuges.
This application note describes a protocol for isolating exosomes from adipose-derived stem cells using Eppendorf’s BioBLU® 0.3c bioreactors and centrifuges. Cells are cultured in bioreactors, then conditioned media is processed via sequential centrifugation steps using a CR22N high-speed centrifuge and CP100NX ultracentrifuge. Exosome purity and yield are enhanced using sucrose cushion methods. The isolated exosomes are characterized using dynamic light scattering, TEM, and CD63 ELISA.
| Centrifuge Model | CR22N |
|---|---|
| Ultracentrifuge Model | CP100NX |
| Rotor Type | P32ST horizontal rotor |
| Exosome Size | 30-150nm |
The tubes must be spun at 100,000 x g for 90 minutes.