Hello,
I have been growing adult neurosphere cultures for a while now by isolating cells myself from the hippocampus and and proliferating the cells as neurospheres for 7days, then plating down and staining. For each culture I do, I have started completely afresh with new cells from new animals. Now though, we want to start passaging the neurospheres so I just have a few questions.
How many passages do people generally maintain neurospheres as floating cultures for? Is it 2-3 passages or 10-11? for example?
My experimental protocol is to treat cells during the neurosphere stage for 7 days, then plate down to stain or differentiate after this. This might seem like an obvious question, but I would assume that I should establish a pool of control (untreated) neurosphere cultures, passage after an appropriate period of growth, then plate down some neurospheres for maintaining the cells and plate down a second set of neurospheres to use for my experiments and treatments?
I am going to set up a culture and examine my cells immunocytochemically after each passage, but I was wondering if anyone could recommend papers/resources that might be good for highlighting the pros/cons and considerations for passaging neurospheres?
Thanks
smr86
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