Human umbilical cord blood cells decrease microglial survival in vitro.
When human umbilical cord blood (HUCB) cells are systemically administered following middle cerebral artery occlusion (MCAO) in rats, they produce a reduction in infarct size resulting in recovery of motor function. Rats receiving HUCB cells have a less severe inflammatory response compared to MCAO stroke rats. The purpose of this study was to determine the interaction between HUCB cells and the main resident immune cells of the brain (microglia) under normoxic and hypoxic conditions in vitro. Primary microglial cultures were incubated for 2 h in no oxygen (95% N, 5% CO(2)) and low glucose (1%) media. Mononuclear HUCB cells were added to half the cultures at the beginning of the hypoxia conditions. Microglial viability was determined using fluorescein diacetate/propidium iodide (FDA/PI) labeling and cytokine expression using ELISA. In some studies, CD11b+ or CD19+ cells isolated from the HUCB mononuclear fraction with magnetic antibody cell sorting (MACS) were used instead of the mononuclear fraction. Co-culturing mononuclear HUCB cells with microglia decreased viability of the microglia during hypoxia. In the microglial monocultures, hypoxia significantly increased release of IL-1beta compared to normoxia, while adding HUCB cells in the hypoxia condition decreased IL-1beta concentrations to the same level as in the normoxia monocultures. Both CD11b+ and CD19+ HUCB cells decreased microglial viability during normoxia and hypoxia. Our data suggest that HUCB cells may produce a soluble factor that decreases viability of microglia.
Center of Excellence for Aging and Brain Repair, University of South Florida, Tampa, Florida 33612, USA.
Lixian
L
Jiang
Tracy
T
Womble
Samuel
S
Saporta
Ning
N
Chen
Cyndy Davis
CD
Sanberg
Paul R
PR
Sanberg
Alison E
AE
Willing
D000818:Animals; D018941:Antigens, CD19 / Q000378:metabolism; D001921:Brain; D039481:CD11b Antigen / Q000378:metabolism; D002450:Cell Communication; D015687:Cell Hypoxia; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism; D005333:Fetus; D005434:Flow Cytometry; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D017628:Microglia / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D018941:Antigens, CD19; D039481:CD11b Antigen; D016207:Cytokines; D005947:Glucose