title: Human placental eXpanded (PLX) mesenchymal-like adherent stromal cells confer neuroprotection to nerve growth factor (NGF)-differentiated PC12 cells exposed to ischemia by secretion of IL-6 and VEGF.
abstract: Mesenchymal stem cells are potent candidates in stroke therapy due to their ability to secrete protective anti-inflammatory cytokines and growth factors. We investigated the neuroprotective effects of human placental mesenchymal-like adherent stromal cells (PLX) using an established ischemic model of nerve growth factor (NGF)-differentiated pheochromocytoma PC12 cells exposed to oxygen and glucose deprivation (OGD) followed by reperfusion. Under optimal conditions, 2 × 10⁵ PLX cells, added in a trans-well system, conferred 30-60% neuroprotection to PC12 cells subjected to ischemic insult. PC12 cell death, measured by LDH release, was reduced by PLX cells or by conditioned medium derived from PLX cells exposed to ischemia, suggesting the active release of factorial components. Since neuroprotection is a prominent function of the cytokine IL-6 and the angiogenic factor VEGF165, we measured their secretion using selective ELISA of the cells under ischemic or normoxic conditions. IL-6 and VEGF165 secretion by co-culture of PC12 and PLX cells was significantly higher under ischemic compared to normoxic conditions. Exogenous supplementation of 10 ng/ml each of IL-6 and VEGF165 to insulted PC12 cells conferred neuroprotection, reminiscent of the neuroprotective effect of PLX cells or their conditioned medium. Growth factors as well as co-culture conditioned medium effects were reduced by 70% and 20% upon pretreatment with 240 ng/ml Semaxanib (anti VEGF165) and/or 400 ng/ml neutralizing anti IL-6 antibody, respectively. Therefore, PLX-induced neuroprotection in ischemic PC12 cells may be partially explained by IL-6 and VEGF165 secretion. These findings may also account for the therapeutic effects seen in clinical trials after treatment with these cells.
authors.affiliation: Pharmacology, School of Pharmacy Institute for Drug Research, The Hebrew University of Jerusalem, P.O. Box 12065, Jerusalem 91120, Israel.
authors.forename: Adi
authors.initials: A
authors.lastname: Lahiani
authors.affiliation: Pluristem Therapeutics Ltd., MATAM Advanced Technology Park #20, Haifa 31905, Israel.
authors.forename: Efrat
authors.initials: E
authors.lastname: Zahavi
authors.affiliation: Pluristem Therapeutics Ltd., MATAM Advanced Technology Park #20, Haifa 31905, Israel.
authors.forename: Nir
authors.initials: N
authors.lastname: Netzer
authors.affiliation: Pluristem Therapeutics Ltd., MATAM Advanced Technology Park #20, Haifa 31905, Israel.
authors.forename: Racheli
authors.initials: R
authors.lastname: Ofir
authors.affiliation: Pluristem Therapeutics Ltd., MATAM Advanced Technology Park #20, Haifa 31905, Israel.
authors.forename: Lena
authors.initials: L
authors.lastname: Pinzur
authors.affiliation: Pluristem Therapeutics Ltd., MATAM Advanced Technology Park #20, Haifa 31905, Israel.
authors.forename: Shani
authors.initials: S
authors.lastname: Raveh
authors.affiliation: Pharmacology, School of Pharmacy Institute for Drug Research, The Hebrew University of Jerusalem, P.O. Box 12065, Jerusalem 91120, Israel.
authors.forename: Hadar
authors.initials: H
authors.lastname: Arien-Zakay
authors.affiliation: Department of Neurobiology, The Weizmann Institute of Science, Rehovot 76100, Israel. Electronic address: ephraim.yavin@googlemail.com.
authors.forename: Ephraim
authors.initials: E
authors.lastname: Yavin
authors.affiliation: Pharmacology, School of Pharmacy Institute for Drug Research, The Hebrew University of Jerusalem, P.O. Box 12065, Jerusalem 91120, Israel.
authors.forename: Philip
authors.initials: P
authors.lastname: Lazarovici
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002452:Cell Count; D016923:Cell Death / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003497:Cyclic N-Oxides / Q000494:pharmacology; D005260:Female; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D015850:Interleukin-6 / Q000378:metabolism*; D007511:Ischemia / Q000473:pathology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D009414:Nerve Growth Factors / Q000494:pharmacology*; D018696:Neuroprotective Agents / Q000378:metabolism*; D016716:PC12 Cells; D010920:Placenta / Q000166:cytology*; D011247:Pregnancy; D011758:Pyrroles / Q000494:pharmacology; D051381:Rats; D013113:Spin Labels; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D017077:Culture Media, Conditioned; D003497:Cyclic N-Oxides; D007211:Indoles; D015850:Interleukin-6; D009414:Nerve Growth Factors; D018696:Neuroprotective Agents; D011758:Pyrroles; D013113:Spin Labels; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C116890:Semaxinib; D007770:L-Lactate Dehydrogenase; C001803:tempol
keywords: Conditioned medium; Human placental mesenchymal-like adherent stromal cells (PLX); IL-6; Ischemia; NGF-differentiated PC12 cells; Neuroprotection; Oxygen and glucose deprivation; Reperfusion; Semaxanib; VEGF