How to differentiate PC12 cells with NGF

Rae Rae
Fri May 27 16:50:10 EST 1994


In article <199405271233.VAA05668 at inetnif.niftyserve.or.jp>
QFH03407 at NIFTYSERVE.OR.JP (=?ISO-2022-JP?B?GyRAQFBETSEhPVM8IxsoSg==?=) writes:
>Dear netters
>I intended to differentiate PC12 cell with NGF. I added NGF in culture
> medium containing 5% horse serum/fetal bovine serum. However, only 
>small part of cells differentiated. How can I overcome this problem ? 
>Is it necessary to change the medium to serum-free before NGF treatment ?
>Please teach me.
>
>======================================================================
>Dr.Toshiharu Ishizuka
>Department of Biochemistry, Chiba University, School of Medicine
>1-8-1, Inohana, Chuo-ku, Chiba, 260 JAPAN
>FAX +81-43-226-2041
>E-mail
>QFH03407 at niftyserve.or.jp
>tishizuk at twics.com
>======================================================================
>

The most important thing to do is to plate the PC12s on a substratum that
promotes neurite outgrowth-  I use a combination of polylysine (added first)
and laminin.  If you don't give them a good substratum the NGF makes them less
adhesive to the plastic and they fall off.  Another problem might be your PC12
cell line- if they've been carried too long and allowed to overgrow before
splitting, then you start selecting for non- responsive cells because they grow
faster than the ones that respond to NGF.  Serum-free medium (ie., supplemented
with N-2) also works better than HS/FCS.  If you want more details E-mail me.

Rae Nishi
nishir at ohsu.edu
Oregon Health Sciences University
Portland, Oregon USA




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