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Am J Physiol Heart Circ Physiol 279: H1434-H1439, 2000;
0363-6135/00 $5.00
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Vol. 279, Issue 3, H1434-H1439, September 2000

SPECIAL COMMUNICATION
Intact endothelial and smooth muscle function in small resistance arteries after 48 h in vessel culture

Steffen-Sebastian Bolz, Susanne Pieperhoff, Cor De Wit, and Ulrich Pohl

Physiologisches Institut, Ludwig Maximilians Universität, D-80336 München, Germany

Long-term culture of resistance vessels allows introduction of molecular biology techniques for use in microvascular research. The aim of the present study was to establish a culture protocol that preserved vascular integrity and function in microvessels for 48 h in culture. Skeletal muscle resistance arteries were excised from the hamster gracilis muscle. Segments were assigned to immediate functional tests or to vessel culture, during which segments were perfused and superfused at a transmural pressure of 45 mmHg with Leibovitz (L15) medium containing 15% fetal calf serum and antibiotics for 48 h. Cultured and freshly isolated vessels showed similar levels of spontaneous tone, myogenic responses, changes in smooth muscle intracellular calcium (Cai2+) (fura 2), and vascular diameter (video microscopy) in response to 0.3 M norepinephrine and similar concentration-response curves for acetylcholine (endothelium dependent, ±Nomega -nitro-L-arginine) and sodium nitroprusside (endothelium independent). Measurements of endothelial Cai2+ revealed similar acetylcholine-induced increases in endothelial Cai2+ in both groups. It is concluded that vascular function can be preserved while maintaining vessels in culture. Thus it is possible to utilize protocols that require long-term treatment.

endothelium-derived hyperpolarizing factor; nitric oxide; calcium measurements; long-term treatment


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