Ultroser G inhibits prostaglandin output by human decidual cells

Dear Sir We have recently measured prostaglandin (PG) production by freshly dispersed human term decidual cells in vitro. To validate our protocol, we investigated the effect of a number of commercially available tissue culture media and media additives on the production of PGF. and PGE. by these cells. Single cell suspensions of human term decidua were prepared by enzymic dispersion and Percoll density centrifugation as previously described. 1’2 The cells were then incubated at 2 x 106 cells/ml for 2 h in various culture media with or without media supplementation. All incubations were carried out under sterile conditions at 37C in a humidified 5% CO2/95% air environment and in the absence of antibiotics; appropriate controls were included. PGF2 and PGE2 output were measured by radioimmunoassay (RIA) of the conditioned medium using antisera raised against the PGs as their methyl oximes. The tissue culture media investigated included RPMI 1640 containing Hepes buffer and 2 mM glutamine (Gibco-Europe, Uxbridge, Middlesex., UK), Minimal Essential Medium Eagle with Earle’s salts (Sigma Chemical Co., Poole, Dorset, UK), Dulbecco’s Modified Eagle’s medium containing 1.0 g/1 glucose (DMEM; Gibco-Europe) and endotoxin-free phosphate buffered saline (PBS; Sigma); medium additives included 10% heat-inactivated normal human serum (NHS; Blood Transfusion Service, John Radcliffe Hospital, Oxford, UK), 0.25% bovine serum albumin (BSA [fraction V]; Sigma) or a serum substitute, Ultroser G, added to a final cortcentration of 2% as recommended (GibcoEurope). PG output by cells cultured in RPMI, DMEM and PBS was similar. Addition of either 10% NHS or 0.25% BSA to any of these culture media had no effect on PG output; however, the addition of 2%