title: Stem cells decreased neuronal cell death after hypoxic stress in primary fetal rat neurons in vitro.
abstract: To explore stem cell-mediated neuronal protection through extracellular signaling pathways by transplanted stem cells, we sought to identify potential candidate molecules responsible for neuronal protection using an in vitro coculture system. Primary fetal rat hippocampal neurons underwent  hypoxia (≤1% oxygen) for 96 h nad then were returned to a normoxic condition. The study group then received rat umbilical cord matrix-derived stem cells, while the control group received fresh media only. The experimental group showed decreased neuronal apoptosis compared to the control  group [44.5 ± 1.6% vs. 71.0 ± 4.2% (mean ± SD, p = 0.0005) on day 5] and higher neuronal survival (4.9 ± 1.2 cells/100× field vs. 2.2 ± 0.3, p = 0.02 on day 5). Among 90 proteins evaluated using a protein array, stem cell coculture media  showed increased protein secretion of TIMP-1 (5.61-fold), TIMP-2 (4.88), CNTF-Rα (3.42), activin A (2.20), fractalkine (2.04), CCR4 (2.02), and decreased secretion in MIP-2 (0.30-fold), AMPK α1 (0.43), TROY (0.48), and TIMP-3 (0.50). This study demonstrated that coculturing stem  cells with primary neurons in vitro decreased neuronal cell death after hypoxia with significantly altered protein secretion. The results suggest that stem cells may offer neuronal protection through extracellular signaling.
authors.affiliation: Department of Anesthesiology, The McGowan Institute for Regenerative Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA. sakait@upmc.edu
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Sakai
authors.forename: Yan
authors.initials: Y
authors.lastname: Xu
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism; D028341:Activins / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D015687:Cell Hypoxia; D002478:Cells, Cultured; D054428:Chemokine CX3CL1 / Q000378:metabolism; D054426:Chemokine CXCL2 / Q000378:metabolism; D053739:Ciliary Neurotrophic Factor Receptor alpha Subunit / Q000096:biosynthesis; D018920:Coculture Techniques; D005260:Female; D005333:Fetus; D006624:Hippocampus / Q000166:cytology / Q000196:embryology; D009474:Neurons / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D054398:Receptors, CCR4 / Q000378:metabolism; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism; D015398:Signal Transduction; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000502:physiology*; D013312:Stress, Physiological; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000378:metabolism; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000378:metabolism; D014470:Umbilical Cord / Q000166:cytology
chemical_list: C515356:Ccr4 protein, rat; D054428:Chemokine CX3CL1; D054426:Chemokine CXCL2; D053739:Ciliary Neurotrophic Factor Receptor alpha Subunit; C513296:Cxcl2 protein, rat; D054398:Receptors, CCR4; D018124:Receptors, Tumor Necrosis Factor; D019715:Tissue Inhibitor of Metalloproteinase-1; D019717:Tissue Inhibitor of Metalloproteinase-3; C422405:activin A; D028341:Activins; D019716:Tissue Inhibitor of Metalloproteinase-2; D055372:AMP-Activated Protein Kinases; C568305:Prkaa1 protein, rat