neuroblastoma stem cells - problems with neurospheres (Sep/28/2007 )
I'm new in this forum and need your help urgently!
I've some problems with my cancer stem cells and because I'm the only one of my lab group who works with this stuff, there's nearly nobody I can talk to. so please help me!!!
I've cancer stem cell lines from two different patients (but it's the same tumour) and they totally behave different although I use the same medium and so on... one of them forms neurospheres very good but the other one makes trouble. the first two passages it forms neurospheres too, but after the third passage more and more neurospheres attach the bottom of the flasks/6-well-plates. the last passage is now 6 days left and today all neurospheres are attached and the cells differentiate like hell. I really don't know what's the problem. are they to old now? but it was only the third passage and I need many many cells because I will do some xenograft stuff with them.
I use NeurobasalA medium with B27-supplement, Heparin, L-glutamine, EGF and bFGF and pen/strep. This should be ok, because it's a neuroblastoma and it works in the other "cellline" very well. And I feed them every second day.
and does anybody know, if I can use them, although they differentiate or do they loose than there stem cell character and are worthless now?
which stem cell markers do you use to detect the real cancer stem cells?
I really appreciate every advice you can give, so thanks...
We are having a similar thing with neurospheres from glioblastomas. And, I have read this in several different publications. Tumors from different patients (althoguh they are diagnosed as the same type) have different neurosphering ability.
If I were you, I would not expect it to be a mistake. Rather, I would use it as a data. As you say, the other patients tumor is a positive control, in this case.