maximizing neuroblastoma adherence to plates

[Sean Patterson]

>Hey,
>
>   We work with N1E-115 cells in our lab.  They seem to stick well to the
>bottom of your typical polystyrene tissue culture dishes (Falcon, Corning)
>when they are undifferentiated.  However, following differentiation, they
>really don't stick very well to anything it seems.  We've tried coating
>the plates with collagen, but that didn't help at all.  We're going to try
>poly-L-lysine next, but after that we're pretty much out of ideas.
>
>   The ultimate goal is to do IHC without the differentiated N1E-115 cells
>sloughing off of the plates.  Does anyone out there have a trick up their
>sleeve they'd be willing to share?
>
>
>Contact,
>
>Jon Roth
>
>jdroth at vet.upenn.edu

Heya Jon,

	I worked with the N1E-115's and a derivative cell line for several
years, and also found problems with the adhesion on differentiation. I
temporarily solved the problem using a standard poly-lysine coat, followed
by an incubation of 1 hour at 37C with foetal calf serum. The plates were
washed and cells plated immediately. The problem with this technique was
that after several days in differentiating medium with DMSO, there was a
tendency for the "pseudo-ECM" to peel off, and I would get lovely
differentiated cells floating on the medium surface with their processes
and all.

	What finally worked for me was the product Cell-Tak from
Collaborative Research (no relation, etc.) - a polyphenolic protein that
simply acts as a molecular glue for membranes. Never had any problems after
that.

Good luck

Sean


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