TY - JOUR
T1 - The brain slice chamber, a novel variation of the Boyden Chamber Assay, allows time-dependent quantification of glioma invasion into mammalian brain in vitro
AU - Schichor, Christian
AU - Kerkau, Siglinde
AU - Visted, Theresa
AU - Martini, Rudolf
AU - Bjerkvig, Rolf
AU - Tonn, Jörg Christian
AU - Goldbrunner, Roland
PY - 2005/5
Y1 - 2005/5
N2 - Glioma cell invasion occurs in a complex micromilieu consisting of neural and glial cells, myelinated fiber tracts, blood vessels and extracellular matrix proteins. The present work describes the brain slice chamber (BSC) as a novel experimental model for assessing invasion of glioma cells into adult mammalian white and gray matter on the basis of the well known Boyden chamber system. As a matrix for invasive tumor cells we used freshly prepared brain tissue from adult pigs. The tissue was sectioned into 40 μm slices that were mechanically fixed to a millipore filter. The neural structures and the three-dimensional architecture of the slice was preserved as verified by immunohistochemistry, light- and electron microscopy. Human U-373 and U87 astrocytoma cells stably transfected with green fluorescent protein (GFP) were assessed for their invasiveness into the brain-slices during a 24 h period. Invasion of U-87 GFP cells was quantified at different time intervals by confocal laser scanning microscopy showing more intense invasion into white compared to gray matter. Two cytostatics (vincristin and paclitaxel) which both are known to affect the cytoskeleton, inhibited glioma cell invasion in a dose dependent manner, which makes the presented model system suitable for functional experiments. In conclusion, the BSC represents a valid and rapid experimental model that may be used to describe the invasive behavior of glioma cells within the preserved three-dimensional structure of mammalian brain tissue in vitro.
AB - Glioma cell invasion occurs in a complex micromilieu consisting of neural and glial cells, myelinated fiber tracts, blood vessels and extracellular matrix proteins. The present work describes the brain slice chamber (BSC) as a novel experimental model for assessing invasion of glioma cells into adult mammalian white and gray matter on the basis of the well known Boyden chamber system. As a matrix for invasive tumor cells we used freshly prepared brain tissue from adult pigs. The tissue was sectioned into 40 μm slices that were mechanically fixed to a millipore filter. The neural structures and the three-dimensional architecture of the slice was preserved as verified by immunohistochemistry, light- and electron microscopy. Human U-373 and U87 astrocytoma cells stably transfected with green fluorescent protein (GFP) were assessed for their invasiveness into the brain-slices during a 24 h period. Invasion of U-87 GFP cells was quantified at different time intervals by confocal laser scanning microscopy showing more intense invasion into white compared to gray matter. Two cytostatics (vincristin and paclitaxel) which both are known to affect the cytoskeleton, inhibited glioma cell invasion in a dose dependent manner, which makes the presented model system suitable for functional experiments. In conclusion, the BSC represents a valid and rapid experimental model that may be used to describe the invasive behavior of glioma cells within the preserved three-dimensional structure of mammalian brain tissue in vitro.
KW - Glioblastoma
KW - Glioma
KW - In vitro
KW - Invasion
KW - Model
KW - Paclitaxel; vincristin
UR - http://www.scopus.com/inward/record.url?scp=20444443203&partnerID=8YFLogxK
U2 - 10.1007/s11060-004-3341-3
DO - 10.1007/s11060-004-3341-3
M3 - Article
C2 - 15933811
AN - SCOPUS:20444443203
SN - 0167-594X
VL - 73
SP - 9
EP - 18
JO - Journal of Neuro-Oncology
JF - Journal of Neuro-Oncology
IS - 1
ER -