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a Hearing Research Lab,
b Molecular and Structural Neurobiology and Gene Therapy Lab, and
c Department of Pathology and Anatomical Sciences, SUNY University at Buffalo, Buffalo, New York, USA;
d NeuroRepair Department, Medical Research Center, Warsaw, Poland
Key Words. Neural stem cells • Human umbilical cord blood • Neurotransmitter receptors • Patch clamp • Inward rectifying potassium current • Outward rectifying potassium current
Correspondence: Michal K. Stachowiak, Ph.D., Department of Pathology and Anatomical Sciences, 206A Farber Hall, SUNY University at Buffalo, Buffalo, New York 14214, USA. Telephone: 716-829-3540; Fax: 716-829-2911; e-mail: mks4{at}buffalo.edu
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),
-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 (IK+). Kir was present on most HUCB-NSCs and HUCB-NSCDs, whereas IK+ 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.
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