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Am J Physiol Cell Physiol 285: C1101-C1108, 2003. First published July 23, 2003; doi:10.1152/ajpcell.00050.2003
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EXTRACELLULAR MATRIX, CELL INTERACTIONS

Fluid pressure in human dermal fibroblast aggregates measured with micropipettes

L. E. B. Stuhr,1 A. Reith,1 S. Lepsøe,1 R. Myklebust,2 H. Wiig,1 and R. K. Reed1

1Department of Physiology and 2Department of Anatomy and Cell Biology, University of Bergen, N-5009 Bergen, Norway

Submitted 5 February 2003 ; accepted in final form 2 July 2003

Previous studies indicated that connective tissue cells in dermis are involved in control of interstitial fluid pressure (Pif). We wanted to develop and characterize an in vitro model representative of loose connective tissue to study dynamic changes in fluid pressure (Pf) over a time course of a few minutes. Pf was measured with micropipettes in human dermal fibroblast cell aggregates of varying size (<100- and >100-µm diameter) and age (days 1-4) kept at different temperatures (~15, 25, and 35°C). Pressures were measured at different depths of micropipette penetration and after treatment with prostaglandin E1 isopropyl ester (PGE1), latanoprost (PGF2{alpha}), and ouabain. Pf was positive (more than +2 mmHg) during control conditions and increased with increasing aggregate size (day 2), age (day 4 vs. day 1), temperature, and depth of micropipette penetration. Pf decreased from 2.9 to 2.0 mmHg during the first 10 min after application of 10 µl of 1 mM PGE1 (P < 0.001). Pf increased from 3.0 to 4.8 mmHg (P < 0.01) after administration of 10 µl of 1.4 µM ouabain and from 3.1 to 4.4 mmHg after addition of 5 µl of 1.42 mM PGF2{alpha} (P > 0.05). In conclusion, we have developed and validated a new in vitro method for studying fluid pressure in loose connective tissue elements with the advantage of allowing reliable and rapid screening of substances that have a potential to modify Pf and studying in more detail specific cell types involved in control of Pf. This study also provides evidence that fibroblasts in the connective tissue can actively modulate Pf.

micropuncture; prostaglandin E1; prostaglandin F2{alpha}; ouabain; integrins


Address for reprint requests and other correspondence: L. E. B. Stuhr, Dept. of Physiology, Univ. of Bergen, Jonas Liesv. 91, N-5009 Bergen (E-mail: linda.stuhr{at}fys.uib.no).






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