Bone marrow mononuclear cells protect neurons and modulate microglia in cell culture models of ischemic stroke.
Although several studies have provided evidence for the therapeutic potential of bone marrow-derived mononuclear cells (MNCs) in animal models of stroke, the mechanisms underlying their benefits remain largely unknown. We have determined the neuroprotective potential of MNCs in primary neuronal cultures exposed to various injuries in vitro. Cortical neurons in culture were exposed to oxygen-glucose deprivation, hypoxia, or hydrogen peroxide, and cell death was assayed by MTT, caspase-3 activation or TUNEL labelling at 24 hrs. Cultures were randomized to cotreatment with MNC-derived supernatants or media before injury exposure. In separate experiments, macrophage or microglial cultures were exposed to lipopolypolysacharide (LPS) in the presence and absence of MNC-derived supernatants. Neuronal cultures were then exposed to conditioned media derived from activated macrophages or microglia. Cytokines from the supernantants of MNC cultures exposed to normoxia or hypoxia were also estimated by enzyme-linked immunosorbant assay (ELISA). MNC-derived supernatants attenuated neuronal death induced by OGD, hypoxia, hydrogen peroxide, and conditioned macrophage/microglial media and contain a number of trophic factors, including interleukin-10, insulin-like growth factor-1, vascular endothelial growth factor, and stromal cell-derived factor-1. MNCs provide broad neuroprotection against a variety of injuries relevant to stroke.
Sharma
Sushil
S
Department of Neurology, University of Texas Medical School at Houston, Houston, Texas 77030, USA.
Yang
Bing
B
Strong
Roger
R
Xi
XiaoPei
X
Brenneman
Miranda
M
Grotta
James C
JC
Aronowski
Jaroslaw
J
Savitz
Sean I
SI
D000704:Analysis of Variance; D000818:Animals; D001854:Bone Marrow Cells / Q000737:chemistry / Q000502:physiology*; D053148:Caspase 3 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016207:Cytokines / Q000378:metabolism; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D005947:Glucose / Q000172:deficiency; D000860:Hypoxia; D020287:In Situ Nick-End Labeling / Q000379:methods; D017628:Microglia / Q000187:drug effects / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D018696:Neuroprotective Agents / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley
D017077:Culture Media, Conditioned; D016207:Cytokines; D018696:Neuroprotective Agents; D016291:Dizocilpine Maleate; D053148:Caspase 3; D005947:Glucose