International Journal of Cell Cloning, Vol 10, 352-358, Copyright © 1992 by AlphaMed Press
Cloning of human tumor cell lines in porous glass capillary tubes: a further development of the human tumor stem cell assay
H Weisser, R Schnabel, P Langer and B Lathan
Institute of Clinical Chemistry and Laboratory Medicine, University Clinic Bergmannsheil, Bochum, Germany.
The conventional human tumor stem cell assay for cloning tumor cells for
drug sensitivity testing is limited by its inability to test drug
combinations. In an attempt to overcome this limitation, we cloned tumor
cell lines within porous glass capillary tubes. In contrast to plastic
porous tubes, the porous glass membranes were transparent, and colony
formation could be judged on an inverted microscope. Human as well as
animal cell lines showed sufficient colony growth. Colonies formed within
these porous tubes were homogeneously distributed, and their morphology was
similar to those formed in the common stem cell assay. Cloning efficiency
and colony size depended on the mean pore diameter of the glass membrane,
with best colony growth within tubes with a pore diameter ranging from 8.5
nm to 14 nm. A linear relationship between number of cells seeded and
number of grown colonies could be demonstrated for the cell lines MDA-231
and Colo 201. Colony growth achieved within porous glass capillary tubes is
comparable to that achieved in Petri dishes and in nonporous tubes. We
conclude that the porous capillary cloning system meets the basic
suppositions for a quantitative cloning assay. Moreover, the porosity of
the glass membrane offers the possibility of variable perfusion of medium
and drugs. Further investigations will focus on various perfusion
modalities and chemosensitivity testing.
