16043459

pubmed:Citation

7

Fetal cells with the characteristics of neural stem cells (NSCs) can be derived from the nonhematopoietic fraction of human umbilical cord blood (HUCB), expanded as a nonimmortalized cell line (HUCB-NSC), and further differentiated into neuron-like cells (HUCB-NSCD); however, the functional and neuronal properties of these cells are poorly understood. To address this issue, we used whole-cell patch-clamp recordings, gene microarrays, and immunocytochemistry to identify voltage-gated channels and ligand-gated receptors on HUCB-NSCs and HUCB-NSCDs. Gene microarray analysis identified genes for voltage-dependent potassium and sodium channels and the neurotransmitter receptors acetylcholine (ACh), gamma-aminobutyric acid (GABA), glutamate, glycine, 5-hydroxytryptamine (5-HT), and dopamine (DA). Several of these genes (GABA-A, glycine and glutamate receptors, voltage-gated potassium channels, and voltage-gated sodium type XII alpha channels) were not expressed in the HUCB mono-nuclear fraction (HUCB-MC), which served as a starting cell population for HUCB-NSC. HUCB-NSCD acquired neuronal phenotypes and displayed an inward rectifying potassium current (Kir) and an outward rectifying potassium current (I(K+)). Kir was present on most HUCB-NSCs and HUCB-NSCDs, whereas I(K+) was present only on HUCB-NSCDs. Many HUCB-NSCDs were immunopositive for glutamate, glycine, nicotinic ACh, DA, 5-HT, and GABA receptors. Kainic acid (KA), a non-N-methyl-D-asparate (NMDA) glutamate-receptor agonist, induced an inward current in some HUCB-NSCDs. KA, glycine, DA, ACh, GABA, and 5-HT partially blocked Kir through their respective receptors. These results suggest that HUCB-NSCs differentiate toward neuron-like cells, with functional voltage- and ligand-gated channels identified in other neuronal systems.

eng

IM

MEDLINE

1066-5099

Print

NLM

931-45

pubmed-meshheading:16043459-Antigens, CD34, pubmed-meshheading:16043459-Antigens, CD45, pubmed-meshheading:16043459-Cell Differentiation, pubmed-meshheading:16043459-Cell Line, pubmed-meshheading:16043459-Electrophysiology, pubmed-meshheading:16043459-Excitatory Amino Acid Antagonists, pubmed-meshheading:16043459-Fetal Blood, pubmed-meshheading:16043459-Glycine, pubmed-meshheading:16043459-Humans, pubmed-meshheading:16043459-Immunohistochemistry, pubmed-meshheading:16043459-Ions, pubmed-meshheading:16043459-Kainic Acid, pubmed-meshheading:16043459-Ligands, pubmed-meshheading:16043459-N-Methylaspartate, pubmed-meshheading:16043459-Neurons, pubmed-meshheading:16043459-Neurotransmitter Agents, pubmed-meshheading:16043459-Oligonucleotide Array Sequence Analysis, pubmed-meshheading:16043459-Patch-Clamp Techniques, pubmed-meshheading:16043459-Phenotype, pubmed-meshheading:16043459-Potassium, pubmed-meshheading:16043459-Receptors, GABA, pubmed-meshheading:16043459-Receptors, Glutamate, pubmed-meshheading:16043459-Sodium, pubmed-meshheading:16043459-Stem Cells, pubmed-meshheading:16043459-Umbilical Veins

Voltage-sensitive and ligand-gated channels in differentiating neural stem-like cells derived from the nonhematopoietic fraction of human umbilical cord blood.

Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't, Research Support, N.I.H., Extramural