In vitro differentiation of size-sieved stem cells into electrically active neural cells.
Size-sieved stem (SS) cells isolated from human bone marrow and propagated in vitro are a population of cells with consistent marker typing, and can form bone, fat, and cartilage. In this experiment, we demonstrated that SS cells could be induced to differentiate into neural cells under experimental cell culture conditions. Five hours after exposure to antioxidant agents (beta-mercaptoethanol +/- retinoic acid) in serum-free conditions, SS cells expressed the protein for nestin, neuron-specific enolase (NSE), neuron-specific nuclear protein (NeuN), and neuron-specific tubulin-1 (TuJ-1), and the mRNA for NSE and Tau. Immunofluorescence showed that almost all the cells (>98%) expressed NeuN and TuJ-1. After 5 days of beta-mercaptoethanol treatment, the SS cells expressed neurofilament high protein but not mitogen-activated protein-2, glial filament acidic protein, and galactocerebroside. For such long-term-treated cells, voltage-sensitive ionic current could be detected by electrophysiological recording, and the intracellular calcium ion, Ca(2+), concentration can be elevated by high potassium (K(+)) buffer and glutamate. These findings suggest that SS cells may be an alternative source of undifferentiated cells for cell therapy and gene therapy in neural dysfunction.
Hung
Shih-Chieh
SC
Department of Orthopaedics, School of Medicine, National Yang-Ming University, Taipei, Taiwan. hungsc@vghtpe.gov.tw
Cheng
Henrich
H
Pan
Chien-Yuan
CY
Tsai
May J
MJ
Kao
Lung-Sen
LS
Ma
Hsiao-Li
HL
D000970:Antineoplastic Agents / Q000494:pharmacology; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D008564:Membrane Potentials / Q000502:physiology; D008623:Mercaptoethanol / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000235:genetics; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D018408:Patch-Clamp Techniques; D014212:Tretinoin / Q000494:pharmacology
D000970:Antineoplastic Agents; D003470:Culture Media; D009419:Nerve Tissue Proteins; D014212:Tretinoin; D008623:Mercaptoethanol; D002118:Calcium