In article <78074.jaeger at jupiter.ifn-magdeburg.de>, Tino J ger
<Tino.Jaeger at ifn-magdeburg.de> wrote:
>subject: hippocampal slice culture
>>I would like to culture hippocampal slices from adult rats.
>To my knowledge, nobody succeeded in doing this so far.
Nobody has succeeded in doing this because neurons from adult animals are
exquisitely sensitive to the anoxia and trauma that necessarily
accompanies the current slice culturing protocols. I have had excellent
results with hippocampal slice cultures (interface configuration as
described by Stoppini et al.) from rat pups up to P16 or so. These will
survive well for months in culture. Going to animals any older leads to
dramatically decreased viability. If you are willing to try some new
things to try to get this to work, you might think about trying:
1. Replace NaCl with sucrose in your dissection buffer,
2. Culture in the presence of kyurenic acid or other general glutamate
3. Jack up the oxygen concentration in your incubator - I would think
about trying 50% as well as 95% because I would worry about oxygen
toxicity in a chronic exposure.
Also, get the brain cold FAST.
Ultimately,though, I think you may be trying to fight the biology of the
system - might be worth a few pilots but I would not be optimistic about
this working very well. Also, I assume you're comfortable with slice
cultures from young animals - if not, you should definitely get this skill
Just because your doctor has a name
Mark Routbort for your condition doesn't mean he
routbort at neuro.duke.edu knows what it is.