A protocol needed to purify cancer cells from tumor tissue - need help! (Nov/29/2006 )
Hi guys,
I'm a newcomer in this forum,glad to see u, everyone! 
recently my boss appointed cancer stem cells as my new field, but i'm not familiar with it. so, when beginnig my research, I'm first faced a technique problem that how to isolate cancer cells from tumor tissue.
is there anybody can help me? I do greatly appreciate your kindness!! 
Plz make your question clear. Do you want to isolate "Cancer Stem Cells" from the tumor?and if yes which cancer? If required I can give you some references.
hello Calvin ,
sorry,maybe I do not express myself well,I mean that, I don't know how to eliminate some normal human cells (such as normal human leukocytes ,endothelial cells, mesothelial cells and fibroblasts) from tumor tissue,so that I can get tumor cells prospectively. thanks for your advice.
you can attach your references on this board.thx! thx!!
Hi,
The system I know is of the way normal or cancer stem cells have been isolated and grown in culture as floating spheres. The tissue is minced, enzymatically and mechanically made in to single cells, filtered through mesh, by gradient centrifugation fibroblasts are separated, RBCs lysed by RBC lysis buffer, and grown on No-Serum, Non attachment condition so that only stem cells or progenitors capable of surviving anoikis remain and other differentiated cells dye out. One of the best ref I have put here and try searching for 'neurosphere cultures' which will give you a lot of details.
(Sorry for a very long single sentence..)
-Prospective identification of tumorigenic breast cancer cells-
Muhammad Al-Hajj*, Max S. Wicha*, Adalberto Benito-Hernandezdagger , Sean J. Morrison*,Dagger ,§, and Michael F. Clarke
PNAS | April 1, 2003 | vol. 100 | no. 7 | 3983-3988
10.1073/pnas.0530291100
The system I know is of the way normal or cancer stem cells have been isolated and grown in culture as floating spheres. The tissue is minced, enzymatically and mechanically made in to single cells, filtered through mesh, by gradient centrifugation fibroblasts are separated, RBCs lysed by RBC lysis buffer, and grown on No-Serum, Non attachment condition so that only stem cells or progenitors capable of surviving anoikis remain and other differentiated cells dye out. One of the best ref I have put here and try searching for 'neurosphere cultures' which will give you a lot of details.
(Sorry for a very long single sentence..)
-Prospective identification of tumorigenic breast cancer cells-
Muhammad Al-Hajj*, Max S. Wicha*, Adalberto Benito-Hernandezdagger , Sean J. Morrison*,Dagger ,§, and Michael F. Clarke
PNAS | April 1, 2003 | vol. 100 | no. 7 | 3983-3988
10.1073/pnas.0530291100
GREAT!!!
Thank you very much!
