Penstrep penicillin and streptomycin and trypsin

All cell and tissue culture media and supplements were purchased as sterile or were filter sterilised through a 0.20 µM filter. Heat-inactivated (hi) foetal bovine serum (FBS) and hi new born calf serum (NCS) were purchased from Gibco (Paisley, Scotland); hi horse serum (HS) from TCS Biosciences (Corby, England); penstrep (penicillin and streptomycin) and trypsin from Bio Whittaker (Wokingham, England); l-glutamine from BDH (Poole, England), gelatine from Sigma (St. Louis, USA). Glassware, distilled water (dH₂O) and phosphate buffered saline (PBS, Oxoid (Hampshire, England) were autoclaved prior to use. Sterile media were stored at 4 °C and used within 2 months. Plasticware were purchased as sterile from Greiner Bio-one, (Kremsmunster, Austria) unless otherwise stated. EPA purchased from IDS Ltd (Boldon, Tyne & Wear, UK) and Palmitate (Sigma-Aldrich, Poole, UK) were complexed with fatty-acid free bovine serum albumin (BSA) (Sigma-Aldrich, Poole, UK). Recombinant human TNF-α was purchased from Calbiochem (Nottingham, England); TRIZOL reagent (Invitrogen, Life Technologies), propidium iodide (PI) stain from BD Biosciences (San Diego, USA) and creatine kinase assay kits from Catachem (Bridgeport, USA). Recombinant human IGF-I was purchased from GroPep (Adelaide, Australia). BCA™ protein quantitation reagents were obtained from Pierce Chemicals (Chester, England, UK).