title: Pretreatment of therapeutic cells with poly(ADP-ribose) polymerase inhibitor             enhances their efficacy in an in vitro model of cell-based therapy in myocardial             infarct.
abstract: The potential of cell-based therapies in diseases involving ischemia-reperfusion             is greatly hampered by the excessive loss of administered cells in the harsh and             oxidative environment where these cells are supposed to act. Therefore, we investigated             if inhibition of poly(ADP-ribose) polymerase (PARP) in the therapeutically added             cells would lead to their increased viability and, subsequently, to an enhanced             effect in an in vitro simulated ischemia-reperfusion (I-R) setting. Ischemic conditions             were simulated by oxygen and glucose deprivation for 160 min using H9c2 rat cardiomyoblast             cells. After 30 min of reperfusion, these cells received 4 types of treatments:             no added cells (I-R model), fluorescently labeled (Vybrant DiD) therapeutic H9c2             cells with vehicle (H9c2) or PARP inhibitor (10 µM or 100 µM PJ34) pretreatment.             We assessed viability (live, apoptotic and necrotic) of both 'postischemic' and             therapeutic cells with flow cytometric analysis using calcein-AM/ethidium homodimer-2             fluorescent staining after 24 h of co-culture. Further measurements on necrosis             and metabolic activity were performed using lactate dehydrogenase (LDH) release             and resazurin based assays. The percentage of surviving therapeutic cells increased             significantly with PARP inhibition (untreated, 52.02±5.01%; 10 µM PJ34, 63.38±4.50%;             100 µM PJ34, 64.99±3.47%). The percentage of necrotic cells decreased in a similar             manner (untreated, 37.23±4.40%; 10 µM PJ34, 26.83±3.49%; 100 µM PJ34, 24.96±2.43%).             Notably, the survival of the cells that suffered I-R injury was also significantly             higher when treated with PARP-inhibited therapeutic cells (I-R model, 36.44±5.05%;             H9c2, 42.81±5.11%; 10 µM PJ34, 52.07±5.80%; 100 µM PJ34, 54.95±5.55%), while necrosis             was inhibited (I-R model, 43.64±4.00%; H9c2, 37.29±4.55%; 10 µM PJ34, 30.18±4.60%;             100 µM PJ34, 25.52±3.47%). In subsequent experiments, PARP inhibition decreased             LDH-release of the observed combined cell population and enhanced the metabolic             activity. Thus, our results suggest that pretreating the therapeutically added             cells with a PARP inhibitor could be beneficial in the setting of cell-based therapies.
authors.affiliation: Institute of Human Physiology and Clinical Experimental Research,                     Semmelweis University, Budapest, Hungary.
authors.forename: Mónika
authors.initials: M
authors.lastname: Szepes
authors.forename: Zsófia
authors.initials: Z
authors.lastname: Janicsek
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Benkő
authors.forename: Attila
authors.initials: A
authors.lastname: Cselenyák
authors.forename: Levente
authors.initials: L
authors.lastname: Kiss
mesh_terms: D000818:Animals; D002460:Cell Line; D018920:Coculture Techniques; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005434:Flow Cytometry; D006836:Hydro-Lyases / Q000032:analysis / Q000378:metabolism; D008315:Malondialdehyde / Q000032:analysis / Q000378:metabolism; D009203:Myocardial Infarction / Q000139:chemically induced / Q000188:drug therapy*; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D051381:Rats; D015427:Reperfusion Injury / Q000188:drug therapy
chemical_list: D004791:Enzyme Inhibitors; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D008315:Malondialdehyde; D006836:Hydro-Lyases; C045581:lactate dehydratase