NGF differentiation of PC12 cells

Charlie Keith keith at zookeeper.zoo.uga.edu
Wed Jan 19 17:40:24 EST 1994


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In article <klosen-180194163226 at 130.104.130.2> klosen at bani.ucl.ac.be (Klosen Paul) writes:
>We are having trouble to differentiate PC12 cells with NGF. So far we have
>tested four different batches of PC12 cells and NGf from 3 different

1. Are your cells rounded or flat? At a frequency of ca 2%, you get
nonresponsive mutants, and they are more adhesive, and may grow
faster,
so they can easily take over a culture.

2. What is your substrate? PC12's grow neurites nicely on adhesive
substrata, or in collagen, but don't do so well on naked TC plastic

3. How long are you waiting? It takes about 3 days to get much more
than little spikes (morphological) from naive cells

4. You can also try to goose them along by adding antimitotics after
1 day of NGF treatment. Neurite outgrowth seems to be inversely
correlated with mitotic index, and it is possible that dividing cells
are taking over.

5. What is your density? If cells are near confluent, they continue to
cycle, and by 4), don't respond well. I generally use 10-20%
confluence.

Charles Keith
University of Georgia
keith at zookeeper.zoo.uga.edu


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1. Are your cells rounded or flat? There are a couple of common

>sources. Has anyone encountered similar problems and how to solve them ????
>
>-- 
>Paul Klosen
>Cell Biology Lab., Catholic University of Louvain
>klosen at bani.ucl.ac.be





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