Immunocytochemical and physiological characterization of a population of cultured human neural precursors.
Human neural precursor cells (HNPC) have recently become commercially available. In an effort to determine the usefulness of these cells for in vitro studies, we have grown cultured HNPCs (cHNPCs) according to the supplier specifications. Here we report our characterization of cHNPCs under nondifferentiating and differentiating growth conditions and make a comparison to primary HNPCs (pHNPCs) obtained at the same developmental time point from a different commercial supplier. We found that under nondifferentiating conditions, cHNPCs expressed nestin, divided rapidly, expressed few markers of differentiated cells, and displayed both 4-aminopyridine (4-AP)-sensitive and delayed-rectifier type K(+) currents. No inward currents were observed. On changing to differentiating culture conditions, a majority of the cells expressed neuronal markers, did not divide, expressed inward and outward time- and voltage-dependent currents, and responded to the application of the neurotransmitters acetylcholine and glutamate. The outward current densities were indistinguishable from those in undifferentiated cells. The inward currents included TTX-sensitive and -resistant Na(+) currents, sustained Ca(2+) currents, and an inwardly rectifying K(+) current. Comparison of the properties of differentiated cells from cHNPCs with neurons obtained from primary fetal cultures (pHNPCs) revealed two major differences: the differentiated cHNPCs did not express embryonic neural cell adhesion molecule (E-NCAM) immunoreactivity but did co-express GFAP immunoreactivity. The co-expression of neuronal and glial markers was likely due to the growth of cells in serum containing medium as the pHNPCs that were never exposed to serum did express E-NCAM and did not co-express glial fibrillary acidic protein (GFAP). The relevance of these results is discussed and compared with results from other neuronal progenitor populations and cultured human neuronal cells.
Piper
D R
DR
Department of Physiology, University of Utah School of Medicine, Salt Lake City 84108, Utah, USA.
Mujtaba
T
T
Rao
M S
MS
Lucero
M T
MT
D015761:4-Aminopyridine / Q000494:pharmacology; D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000963:Antimetabolites / Q000493:pharmacokinetics; D015415:Biomarkers; D001973:Bromodeoxyuridine / Q000493:pharmacokinetics; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D019070:Cell Lineage / Q000502:physiology; D002478:Cells, Cultured; D005333:Fetus / Q000737:chemistry / Q000166:cytology; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000032:analysis; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology; D007700:Kinetics; D008869:Microtubule-Associated Proteins / Q000032:analysis; D009419:Nerve Tissue Proteins; D064231:Nestin; D019006:Neural Cell Adhesion Molecules / Q000032:analysis; D016900:Neurofilament Proteins / Q000032:analysis; D009474:Neurons / Q000737:chemistry* / Q000166:cytology / Q000502:physiology*; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism; D015221:Potassium Channels / Q000502:physiology; D024661:Potassium Channels, Inwardly Rectifying; D015222:Sodium Channels / Q000502:physiology; D013234:Stem Cells / Q000737:chemistry* / Q000166:cytology / Q000502:physiology*; D013779:Tetrodotoxin / Q000494:pharmacology; D014404:Tubulin / Q000032:analysis
D000963:Antimetabolites; D015415:Biomarkers; D005904:Glial Fibrillary Acidic Protein; D007381:Intermediate Filament Proteins; D008869:Microtubule-Associated Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D019006:Neural Cell Adhesion Molecules; D016900:Neurofilament Proteins; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D015222:Sodium Channels; D014404:Tubulin; D013779:Tetrodotoxin; D015761:4-Aminopyridine; D001973:Bromodeoxyuridine; D011188:Potassium; D002118:Calcium