Obtaining cells from human tissue samples - (Nov/13/2006 )
I am looking for a protocol or a trail to lead me to a protocol for obtaining whole cells from frozen human breast tissue samples. I have samples ranging from less than 0.5g to nearly 10g. There are protocols for obtaining cells from live tissue but I do not know how to translate these so they may be used on frozen tissue sections. Any repsonse is appreciated.
I had similar problems with trying to find out how to use frozen tissue samples myself (the one main difference between us was that they were mouse glioma cells). I believe however that perhaps a similar protocol may be used for your cells.
This is was one protocol I was given:
(I don't know if this would work for non tumor tissue samples ... but it might be worth trying ... if you have enough samples)
"Chop the tumor into 1 mm pieces with crossed blades. Wash the
pieces 2-3x by PBS, then transfer the tumor pieces into a 25 cm2 flask. Add 1.5 ml medium containing 40% serum and antibiotics. The tumor pieces should attach to the dish surface in 4 days. Then you should add medium to 5 ml and adjust serum to 20%. Tumor cells should be grown out from the tumor pieces in 1-2 weeks, then you can collect and pass them by normal treatment."
Alternatively I posted a topic with a similar question quite a long time ago ... link
A couple of the replies gave very informative descriptions of procedures that work with frozen tissue samples.
Hope some of this helps
Thanks for the response. I have pathology reports for each sample and the notes read "fresh sample submitted for frozen section". Also, we are storring them in -80C not in our N2. I will definitely have to check this out. Thanks again and I'll keep checking back.