EFFECT OF COLD PRESERVATION ON PULMONARY ARTERIAL SMOOTH-MUSCLE CELLS

The efficacy of preservation fluids on the cytoskeleton and contractil e function of porcine pulmonary arterial smooth muscle (SM) cells duri ng cooling and rewarming was evaluated, using EuroCollins solution (EC ), University of Wisconsin solution (UW), Marshall's solution (MS), an d tissue culture growth medium (GM). Functional studies included passi ve distensibility and contraction to prostaglandin F-2 alpha (PGF(2 al pha)) 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 w ere stained with antibodies to SM alpha-actin, SM myosin, and tubulin. In cooling, all solutions resulted in increased arterial distensibili ty, whereas EC and MS reduced cell wrinkling. With the use of all solu tions, actin cables thinned, myosin filaments dissociated, and microtu bules depolymerized. During rewarming, resistance to imposed tension i ncreased in all arterial rings. After GM, EC, and MS preservation, con traction to PGF(2 alpha) increased. Wrinkling increased and actin-myos in 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 preserva tion and after rewarming. The absence of spontaneous contraction in ce lls cooled in UW may be advantageous.