Cargo packaged into exosomes is tightly linked to the behaviour of the cell releasing them. eg a stressed cell would release exosomes with a different protein and RNA composition as opposed to when it is not stressed. I would like to know that apart from passage number ,cell seeding density and cell culture conditions (ph , temp , % CO2 , humidity) what else should be taken into consideration in order to make the cells behave consistently on sub - culturing so that they release qualitatively uniform exosomes every time. Or what factors affect cell behaviour in vitro.
First I would like to suggest you that please check whether the cells are in same phase. That is all the cells are growing in same rhythm of cycle, what we call as cell synchrony. I donot know the details of the research, like do your exosomes have variable conc. in variable cell phase. So, I guess this fact should be taken into account.
All the best,
Somnath.
Synchronizing the growth by starvation or low concentration FCS containing media.
However, it is not fully possible to maintain them in the same phase. In addition, exsosome release/yield may vary from cell to cell although they are in the same condition.
I do know nothing about exosomes, but like it has been said previously, to synchronize your cells, you should starve them by reducing the concentration of FCS in your medium.
thank you somnath , venkatesh and lauriane for your help . This was a crucial point which i failed to consider.
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