How long should I cultivate cells for exosome isolation? Are exosomes taken up by the same cells during longer cultivations? Thank you!
Every day cells secret and uptake exosmes.
depending cell type, the number of secreted exosomes are different.
I would collect exosomes every day or every two day.
Exosomes are indeed secreted by cells everyday. Our lab isolates exosomes and microvesicles from cells grown as spheroids in serum-free media after 7 days in culture. You can find the exact protocol using this link:
http://www.molecular-cancer.com/content/12/1/62
In regards to your question about exosomes being taken up by other cells, yes, cells in culture do take up exosomes being produced by the same cells in culture. However, cells in a tissue culture flask produce millions of exosomes, so I would not worry about this affecting the overall number you are able to isolate.
Thank you all! I noticed that people isolate exosomes after 48 or 72 hours of cultivation. I was thinking if there is direct proportion between longer time of cell cultivation-more cells present and exosome yield. Maybe not when they are taken back by cells. I isolate exosomes after 7 days. Thank you for article link, I will read it. Does any papers focus on how many exosomes are released and uptaken by cell during time period?
Using uninfected Jurkat and chronically HIV-1 infected J1.1 T-cell lines we found that seeding our cultures to 1*10^6 cells/mL and then harvesting exosomes at day 5 resulted in much higher isolated exosome levels than harvesting at days 1 or 2. A link to the article is below.
http://www.jbc.org/content/288/27/20014.long
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