In article <Pine.SOL.3.91.960202134756.13965A-100000 at acsusun>,
Neil Armstong <s9590046 at acsusun> wrote:
>Does anyone have a experience in cultuing neutrophils in vitro. I want to
>perform a few experiments and don't have the time to optimise the culture
>conditions for myself. I would appreciate it if anyone could help me out.
I know that'd I'd be impressed if you succesfully cultured human
neuts! Well, actually, if you could do it with any neuts or
granulocytes I'd be impressed...
Maybe your talking about LTBMC or such? If your just talking about a
constant, significant source of normal cells, then I'm kinda'
confused.
Um, you know these cells are (mostly) terminally differentiated, and
hair-trigger by design, so getting them to live outside the body for
any extended time, and in a "normal" unactivated state, is virtually
impossible (as far as I know). BUT, I'd love to hear to the contrary.
Anyway, they're so dang easy to get, why even try to culture them?
Unless you're trying to get murine cells, you'd spend as much time in
TC (and certainly expense), than just drawing some blood, doing a
ficoll spin and ACK lysis (among other easy protocols). You know those
pesky mice just don't have too much blood to get at, and most people
would frown on the massive sack necessary for most experiments ;-)
If you are wanting them from smaller animals, well, most people use
bone marrow exudate.
Oh, make sure you have protocol approval, human consents, and try for
some sort of remuneration. That way when you inevitably end up drawing
from yourself and lab mates (although we never do this as it's
possibly dangerous to work with your own cells, and limits what anyone
would qualify as "normal" volunteers), you'll have money to buy the
coffee and orange juice you'll be drinking afterwards!
Best of luck,
Aaron "No, I'm not a junkie" Warnock
--
"Nothing more is needed to destroy a man, than the conviction that his
life's work is useless." -Antonin Artaud
erawtech at leland.stanford.edu (R. Aaron Warnock)
