can anyone recognize these things? - (Oct/25/2007 )

i recently try to do primary culture of glioblastoma with brain tumor specimens
i chop the tissue into small chunks by scissors and forceps (all sterilize)
treated by collagenase at 37℃
add EGF to medium and incubate the tissues in 37℃ incubator
and found somethings i don't know have been growing
can someone help me in recognize these things?

100x http://upload.imgspot.com/u/07/296/22/0806No.1A100x.jpg
200x http://upload.imgspot.com/u/07/296/22/0806No.1A200x.jpg

100x http://upload.imgspot.com/u/07/296/22/0921No.2A1100x.jpg
200x http://upload.imgspot.com/u/07/296/22/0921No.2A1200x.jpg

100x http://upload.imgspot.com/u/07/296/22/0921No.2A2100x.jpg

are these irregular things neurospheres?
if i want to get glioma or glioblastom cells, are these the target?


and as you can see from the pictures,
the medium is quiet turbid,
if there's a way to get rid of those debris?

any reply will be great helps!!
thank you very much!!

i have no experience with the particular cells you are aiming for...but do use a similar technique for primary smooth muscle cell culture...when I get debris (like the "stuff" you have in your background) it indicates contamination to me - but the globular things, sorry I have no idea on that one. if they are adherent i suppose you could just gently wash away the background by doing several media flushes. good luck, hope you get an answer!

my first thought on the turbidity is bacterial infection - i had it recently in my rho zeros.
you can try treating it with antibiotics but the only thing this would give you is experience in the resistant nature of bacteria - if it doesnt look right sometimes your only option is to chuck it and start again

sorry

dom

(we do skin by the way but i doubt its that different)