I used all trans retinoic acid to differentiate neurospheres generated from medulloblastoma cell lines. I observed neuronal cells but not in sufficient amounts. Can anyone suggest how I can get a higher number of neuronal marker positive cells?
if you are generating neurospheres using EGF and bFGF, then withdrawal of these factors and plating the neurospheres on PDL/laminin-coated dishes should give diffferentiated neurons in a few days.
you are right and I am doing the same. But I used All trans retinoic acid for the neuronal differentiation of neurospheres after plating them on poly-orniithin coated plates. I observed neuronal differentiation but the number is not sufficient to store for further studies. So, I need a protocol to get more number of neuronal cells.
I met the same question, when I withdraw the mitogen, and induce the neurosphere into neurons with RA, but only 10% cells can transfer into NF positive cells. So I have to do the primary culture from E17 embryos.
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