Effect of cold preservation on pulmonary arterial smooth muscle cells

Abstract

The efficacy of preservation fluids on the cytoskeleton and contractile function of porcine pulmonary arterial smooth muscle (SM) cells during cooling and rewarming was evaluated, using EuroCollins solution (EC), University of Wisconsin solution (UW), Marshall's solution (MS), and tissue culture growth medium (GM). Functional studies included passive distensibility and contraction to prostaglandin F2a (PGF2a) in arterial rings and wrinkling of silicone membranes by cooled-rewarmed cultured SM cells. Immunofluorescence measurements were made of actin brightness in cooled arterial rings. Cultured SM monolayers were stained with antibodies to SM alpha-actin, SM myosin, and tubulin. In cooling, all solutions resulted in increased arterial distensibility, whereas EC and MS reduced cell wrinkling. With the use of all solutions, actin cables thinned, myosin filaments dissociated, and microtubules depolymerized. During rewarming, resistance to imposed tension increased in all arterial rings. After GM,ED, and MS preservation, contraction to PGF2a increased. Wrinkling increased and actin-myosin cables shortened after GM and EC; after UW, wrinkling decreased and actin-myosin cables thinned. No recovery occurred after MS. Thus the type of preservation solution influenced contractility during preservation and after rewarming. The absence of spontaneous contraction in cells cooled in UW may be advantageous.