Development of a cell culture system to study antibody convection in tumors.
The convective transport of fluid and of a binding antibody through a cultured tumor cell layer was investigated with a mouse melanoma cell line (B16F10) grown on a microporous polycarbonate filter (Snapwell inserts). The inserts were precoated with Matrigel or collagen, or were uncoated. The cell layers were exposed to nominal pressure gradients from 5 to 25 cm H2O, and the volume flux was measured by collecting the effluent volume over time. The rate of convective transport of a binding monoclonal antibody that recognizes the murina transferrin receptor (a-TfR) was investigated at a nominal pressure gradient of 15 cm H2O and compared with that of an isotype matched, nonbinding control. The resistance, R, of the cell layer to fluid flow was quantified as the hydraulic conductivity, Lp (= 1/R); the ability of the cell layer to retard antibody transport was quantified as the reflection coefficient, sigma. The resulting Lp values decreased with increasing cell density, in a manner consistent with Poiseuille flow. Collagen or Matrigel precoating also decreased Lp values, with cells grown on Matrigel providing the greatest resistance. The sigma values were 0.67 (+/-0.08) for the a-TfR antibody and 0.51 (+/-0.06) for the control, indicating that the cell layer acts as a semipermeable barrier to convective transport of antibody that is less permeable to the binding antibody.
DeSilva
B S
BS
Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, USA.
Hendrickson
T L
TL
Topp
E M
EM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D001692:Biological Transport; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D008546:Melanoma, Experimental / Q000276:immunology; D051379:Mice; D008854:Microscopy, Electron; D011312:Pressure; D011990:Receptors, Transferrin / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D011990:Receptors, Transferrin