Voltage-sensitive and ligand-gated channels in differentiating neural stem-like cells derived from the nonhematopoietic fraction of human umbilical cord blood.
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.
Sun
Wei
W
Department of Pathology and Anatomical Sciences, 206A Farber Hall, SUNY University at Buffalo, Buffalo, New York 14214, USA.
Buzanska
Leonora
L
Domanska-Janik
Krystyna
K
Salvi
Richard J
RJ
Stachowiak
Michal K
MK
D018952:Antigens, CD34 / Q000096:biosynthesis; D002454:Cell Differentiation; D002460:Cell Line; D004594:Electrophysiology; D018691:Excitatory Amino Acid Antagonists / Q000378:metabolism; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism*; D005998:Glycine / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry; D007477:Ions; D007608:Kainic Acid / Q000378:metabolism; D017493:Leukocyte Common Antigens / Q000096:biosynthesis; D008024:Ligands; D016202:N-Methylaspartate / Q000378:metabolism; D009474:Neurons / Q000378:metabolism*; D018377:Neurotransmitter Agents / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D018408:Patch-Clamp Techniques; D010641:Phenotype; D011188:Potassium / Q000737:chemistry; D018079:Receptors, GABA / Q000378:metabolism; D017470:Receptors, Glutamate / Q000378:metabolism; D012964:Sodium / Q000737:chemistry; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014471:Umbilical Veins / Q000378:metabolism
D018952:Antigens, CD34; D018691:Excitatory Amino Acid Antagonists; D007477:Ions; D008024:Ligands; D018377:Neurotransmitter Agents; D018079:Receptors, GABA; D017470:Receptors, Glutamate; D016202:N-Methylaspartate; D012964:Sodium; D017493:Leukocyte Common Antigens; D011188:Potassium; D007608:Kainic Acid; D005998:Glycine