Cell column chromatography: a new research tool to quantify cerebral cell volume changes following chemically-induced anoxia/re-oxygenation.
OBJECTIVE
The roles of individual types of cerebral cells in contributing to brain edema are undefined. The objective of this study was to determine the role of cerebral cell-column chromatography in quantifying cell volumes of individual cerebral cell lines, under chemically-induced anoxia/re-oxygenation (A/R).


METHODS
Cerebral endothelial cells (4 experiments) or type II astrocytes (4 experiments) were cultured to confluence on microcarrier beads. A chromatographic cell-column of 1.5 cm height was filled with non-treated cell-covered beads. The column was perfused at 1 ml/min with a balanced perfusate for one hour (Baseline). The perfusate was then switched to that containing 5 mM thioglycolic acid for one hour (Anoxia). Then the column was perfused with the normal perfusate for another two hours (Re-oxygenation). The total free space in the column, reversely reflecting cell volumes, was determined by averaged transit time (TTa) of a non-permeable flow tracer blue dextran. Decreased TTa means that cells swell, and vice versa.


RESULTS
TTa in endothelial cell columns increased with a peak at 60 minutes of re-oxygenation. TTa in astrocyte columns decreased with a nadir at 30 minutes of re-oxygenation.


CONCLUSIONS
Cell column chromatography can be used to determine the cerebral cell volume changes following chemically-induced anoxia/re-oxygenation.
Xiao
F
F
Department of Emergency Medicine, Louisiana State University Health Science Center, Shreveport 71130, USA. fxiao@lsuhsc.edu
Pardue
S
S
Nash
T
T
Arnold
T C
TC
Alexander
J S
JS
Carden
D L
DL
Turnage
R
R
Jawahar
A
A
Conrad
S A
SA
D001929:Brain Edema / Q000139:chemically induced / Q000473:pathology*; D002452:Cell Count / Q000379:methods; D002469:Cell Separation / Q000379:methods*; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured; D002845:Chromatography / Q000379:methods*; D042783:Endothelial Cells / Q000187:drug effects / Q000473:pathology*; D006801:Humans; D015427:Reperfusion Injury / Q000139:chemically induced / Q000473:pathology*; D013864:Thioglycolates
D013864:Thioglycolates