RT-PCR with cultured neuronal cells

Wolfgang Schechinger hubahopp at gmx.de
Fri Jun 11 13:30:16 EST 2004

Dear Patrick, 

is there a possibility to kill the feeder cells by irradiation before you
seed your neurons? Maybe you can enrich your neurons by MACS as an

Best Reagrds,


At 17:17 11.06.2004, Patrick Wefstaedt wrote:
>I´m trying to investigate the effects of  Neurotrophic Factors on
>neuronal cells from the inner ear (SGC). The cells are kept in culture
>for 72h and were harvested afterwards.  In order to determin the
>expression levels of various receptors RT-PCR is carried out. As
>qualitative proof everything is working fine. My question to you is
>wether there is a possibility to try out semiquantitative experiments,
>too. The problem is that the RNA from the neuronal cells is decreasing
>(because of decreasing number of cells) while the total RNA amount is
>increasing as a reason of fibroblast growth. This means that
>normalization against a “normal” housekeeping gene e.g.
>GAPDH is not possible. Does anybody know about a gene which could be
>taken as a “neuronal” housekeeping gene and which is not
>regulated after exposition to growth factrors or serum? Do you have
>other ideas?
>Thanks a lot!
>Immer auf dem aktuellen Stand mit den Newsgroups von freenet.de:
Wolfgang Schechinger


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