We are going to isolate extracellular vesicles from HL60 cell culture, but we have to culture these cell with FBS. FBS contains Extracellular vesicle which can cause interference.
Do you have a solution for this issue?
Hello Mehrdad Izadirad
You can use chemically defined medium which contain components whose chemical compositions and structures are known, and their chemical species are specified. Such a medium often contains a mixture of recombinant components like growth factors, cytokines, hormones and other proteins. But you will have to adapt HeLa cells to grow in such chemically defined medium.
I would suggest that instead of trying a serum replacement you can always try reducing the amount of serum. Try the least concentration of FBS in which HeLa cells show optimum growth without much difference. You can go down from
10% FBS to 7-8% FBS.
All the Best.
There are several options, depending on how 'robust' your cells are.
- Use of serum free media - this can be done in a hybrid strategy in which you culture cells to 70-80% confluency, wash and then add serum free media for EV collection.
-Use of 1% FBS to limit FBS-EV presence (suggested by Thery et al, but I've never seen anyone use this option).
-Use of a protocol to deplete your FBS of EVs; most commonly ultracentrifugation for 18-hours, 120,000g, dilution is optimal, but not always used.
Obviously each has their own pros and cons, but our cells only retain their cellular properties using EV depleted FBS.
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