Effect of short-term organoid culture on the pharmaco-mechanical properties of rat extra- and intrapulmonary arteries.

1 Organoid cultured explants from differentiated tissues have gained renewed interest in the undertaking of physiological and pharmacological studies. In the work herein, we examined the pharmaco-mechanical properties of an in vitro model consisting of organoid cultured rings derived from rat extra- and intrapulmonary arteries, over a period of 4 days in culture. 2 Mechanical changes were quantified using isometric tension measurements on both fresh and cultured pulmonary arterial tissues, with experiments performed in the presence or absence of 10% foetal calf serum. Conventional histochemical and immunofluorescent stainings were also performed to assess tissue structure integrity and apoptosis. 3 The explants developed spontaneous rhythmic contractions (SRC) in approximately half of the vessels. SRC amplitude and time course were modified by conditions and agents acting on membrane potential (high-potassium solutions--levcromakalim, a potassium channel opener), while nitrendipine, an L-type calcium channel blocker, suppressed SRC. 4 Cultured explants also developed a hyper-reactivity to high potassium challenges (10-40 mM). Whereas contraction to serotonin (5-HT) was enhanced in intrapulmonary arteries, contraction to endothelin-1 remained unchanged after 4 days of culture. Serum did not alter contractile properties during the culture period. 5 Endothelial-dependent relaxation was maintained in response to A23187 500 microM, but was abolished in response to 10 microM carbamylcholine. 6 Histological and immuno-histological analyses revealed the absence of hypertrophied vascular wall or apoptosis. 7 In conclusion, the contractile phenotype as well as tissue structure integrity of organoid explants remain essentially intact during short-term culture, making this model suitable for pharmaco-genomic studies.