Coat the plates with Matrigel prior to plating the cells. It worked for
SY5Y and PC12 cells. Make diluted Matrigel solution (1:20 with water),
vortex until totally dissolved, totally cover plate well with solution,
allow to sit for 3hr. under hood, aspirate off all solution, allow to dry
overnite in hood. With above cells, it takes 5-10 days but extensive
neurite outgrowth is very evident because RA slows cell proliferation down
radically.
Marc C. Brande, M.S. SD3D Email List:3D Imaging
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On 7 Dec 1994, Christopher B. Chambers wrote:
> I have recently begun working with 2 neuroblastoma cell lines (LA-N-2 and
> SK-N-MC) and have been attempting to differentiate them with retinoic
> acid. After approx. 1 week of treating the cells, I have seen no
> alterations in growth of the cells nor have I detected any neurite
> outgrowth. I dissolved the retinoic acid (RA) in 100% ethanol and treat the
> cells every day with 10 microM RA or 0.1% ethanol vol/vol (control). Any
> suggestions on modifications to my protocol or alternative protocols
> would be greatly appreciated.
>> Thanks, Christopher Chambers
>
