Dear Letizia and Bill,
we had also severe problems with the neurons from rat embryos.
This changed a lot, when we were testing Neurobasal A medium from
Invitrogen (please remember, I do not intend to do a PR show here,
it's only help!!!).
We tried many things, like low oxygen, low osmolarity, sandwich
coverslip culture, astrocyte feeding layer (which seems to be really
great, but we have problems with the layer...), different media from
many companies and suffered through all this, sometimes great
cultures, sometimes only dead cells in the well / dish.
After all we started using this literature-source as background:
Acute Effects of Ethanol on Kainate Receptors with Different Subunit
Compositions, Valenzuela, Cardoso2, JPET March 1, 1999 vol. 288
no. 3 1199-1206
http://jpet.aspetjournals.org/content/288/3/1199.abstract
This has nothing to do with our research, but is providing very good
and stable culture results.
I can provide you our actual protocol, if you want, but this should be
done out of the list.
But remember, that the pattern of morphology/ type of the neurons in
this culture is quite different from embryonal cultures.
Best regards
Thomas
Quoting neur-sci-request from oat.bio.indiana.edu,
Neur-sci Digest, Vol 54, Issue 6 (Thu 19 Nov 2009 18:05:41 CET):
> 1. Re: neuronal cultures (Bill)
>>> ----------------------------------------------------------------------
>> Message: 1
> Date: Wed, 18 Nov 2009 16:42:53 -0800 (PST)
> From: Bill <connelly.bill from gmail.com>
> Subject: [Neuroscience] Re: neuronal cultures
> To: neur-sci from net.bio.net> Message-ID:
> <3c551ba0-e7fd-4293-9d66-5298f04bd381 from b25g2000prb.googlegroups.com>
> Content-Type: text/plain; charset=ISO-8859-1
>> Adult cortical neuronal cell culture? Yes, it is difficult to get any
> live neurons in your cell culture. We always work with either E16-17
> or P1-3. For adult neuronal culture you need to do all kinds of
> dissociation, gradient separation, and neurotrophic stuff (at least in
> my experience). Do some searches for adult neuronal culture methods
> and I think you'll see what I mean.
>>> On Nov 14, 9:44Â am, letizia polito <letizia.pol... from gmail.com> wrote:
>> Hi,
>> does anyone work with hippocampal or cortical adult neurons? What are the
>> main problems vs embrional or postnatal cultures? It is difficult to have a
>> neuronal preparation not contaminated by microglia?
>> Thank you everyone.
>>>> ------------------------------
>> _______________________________________________
> Neur-sci mailing list
>Neur-sci from net.bio.net>http://www.bio.net/biomail/listinfo/neur-sci>> End of Neur-sci Digest, Vol 54, Issue 6
> ***************************************
>
--
~~~~~~~~~~~~
Thomas Breitenbach, PostDoc
Dept. of Physiology/ Chemistry; University of Aarhus
Ole Worms Allé00 160, room 115/ Langelandsgade 140, room 1511-221
DK-8000 Aarhus C
phone: +45 8942 2797
fax: +45 8619 6199
mobil: +45 2298 7186
~~~~~~~~~~~~
