Perfusion of single tumor microvessels: application to vascular permeability measurement.
OBJECTIVE
To develop a new method for determining the relative importance of convection versus diffusion in macromolecular transport across tumor microvessel walls.


METHODS
The human colon adenocarcinoma LS174T was transplanted in the dorsal skinfold chamber in a severe combined immunodeficient (SCID) mouse. The vasculature at the tumor surface was exposed by carefully removing the glass window of the chamber. A tumor microvessel was randomly selected, which was approximately 20-40 microns in diameter, embedded in the connective tissue 10-12 microns below the surface of the tumor. The vessel was cannulated with a micropipette and perfused with fluorescein isothiocyanate (FITC)-labeled bovine serum albumin (BSA) at different perfusion pressures. The fluorescence intensity was recorded on videotapes via a video system attached to the fluorescence microscope for offline analysis. The apparent vascular permeability was determined based on the time-dependence of fluorescence intensity and the vessel diameter.


RESULTS
The apparent vascular permeability of single vessels to FITC-labeled BSA was quantified at perfusion pressures of 20-45 cmH2O. The pressure dependence of vascular permeability in LS174T tumors was heterogeneous. On average, there was no correlation between the apparent vascular permeability and the perfusion pressure in the range of 20-35 cmH2O (p = 0.73), even though the apparent permeability increased significantly when the pressure was increased from 20 to 45 cmH2O (p = 0.008).


CONCLUSIONS
These results indicate that convection in the transvascular transport of albumin is not significant in non-peripheral regions of solid tumors in which the pressure difference across the vessel wall is small or even negligible. In addition to the permeability studies, this preparation can be used to study cell-cell interactions in single tumor vessels under defined flow conditions.
Lichtenbeld
H C
HC
Department of Radiation Oncology, Massachusetts General Hospital, Harvard Medical School, Boston 02114, USA.
Yuan
F
F
Michel
C C
CC
Jain
R K
RK
D000230:Adenocarcinoma / Q000098:blood supply*; D000818:Animals; D001794:Blood Pressure; D002199:Capillary Permeability / Q000502:physiology*; D002404:Catheterization; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D006801:Humans; D007700:Kinetics; D051379:Mice; D016513:Mice, SCID; D008833:Microcirculation / Q000502:physiology*; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D009368:Neoplasm Transplantation; D010477:Perfusion; D012710:Serum Albumin, Bovine / Q000493:pharmacokinetics; D014183:Transplantation, Heterologous
C039217:fluorescein isothiocyanate bovine serum albumin; D012710:Serum Albumin, Bovine; D016650:Fluorescein-5-isothiocyanate