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Am J Physiol Heart Circ Physiol (September 12, 2002). doi:10.1152/ajpheart.00327.2002
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Articles in PresS, published online ahead of print September 12, 2002
Am J Physiol Heart Circ Physiol, 10.1152/ajpheart.00327.2002
Submitted on April 10, 2002
Accepted on September 10, 2002

Isolation of interstitial fluid from rat mammary tumors by a centrifugation method

Helge Wiig1*, Knut Aukland1, and Olav Tenstad1

1 Department of Physiology, University of Bergen, Bergen, Norway

* To whom correspondence should be addressed. E-mail: helge.wiig{at}fys.uib.no.

Access to interstitial fluid is of fundamental importance to understand tumor transcapillary fluid balance, including the distribution of probes and therapeutic agents. Tumors were induced by gavage of di-methyl-benz-anthracene to rats, and fluid was isolated after anesthesia by exposing tissue to consecutive centrifugations from 27 to 6800 G. The observed 51Cr-EDTA (extracellular tracer) tissue fluid to plasma ratio obtained from whole tumor or from superficial tumor tissue by centrifugation at 27-424 G was not significantly different from 1.0 (0.92-0.99), suggesting an extracellular origin only. However, fluid collected from excised central tumor had a significantly lower ratio (0.66-0.77) for all imposed G-forces, suggesting dilution by fluid deriving from a space unavailable for 51Cr-EDTA. The colloid osmotic pressure in tumor fluid was generally higher than in fluid isolated from subcutis, attributable to less selective capillaries and impaired lymphatic drainage in tumors. HPLC analysis of tumor fluid showed that low molecular weight macromolecules not present in arterial plasma were present in tumor fluid obtained by centrifugation and in venous blood draining the tumor, most likely representing proteins derived from tumor cells. We conclude that low speed centrifugation may be a simple and reliable method to isolate interstitial fluid from tumors.




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