Highly efficient generation of glutamatergic/cholinergic NT2-derived postmitotic human neurons by short-term treatment with the nucleoside analogue cytosine β- d -arabinofuranoside
| Autor: | Joan Sallés, Xabier Aretxabala, Imanol González-Burguera, Sergio Barrondo, Ana Ricobaraza, Maider López de Jesús, Gontzal García del Caño |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Tyrosine 3-Monooxygenase Neurogenesis Cellular differentiation Blotting Western Retinoic acid Biology Choline O-Acetyltransferase 03 medical and health sciences Glutamatergic chemistry.chemical_compound 0302 clinical medicine Cytosine β-d-arabinofuranoside Cell Line Tumor Animals Humans Cholinergic neuron lcsh:QH301-705.5 Cell Proliferation Medicine(all) Glutamate Decarboxylase Cytarabine Brain Cell Differentiation Cell Biology General Medicine Postmitotic human neurons Cholinergic Neurons Rats Pluripotent NT2 cells Cell biology 030104 developmental biology Microscopy Fluorescence Neuronal differentiation lcsh:Biology (General) Biochemistry chemistry Cell culture Vesicular Glutamate Transport Protein 1 Cholinergic Neurotransmitter phenotype Nucleoside 030217 neurology & neurosurgery Developmental Biology |
| Zdroj: | Stem Cell Research, Vol 16, Iss 2, Pp 541-551 (2016) |
| ISSN: | 1873-5061 |
| Popis: | The human NTERA2/D1 (NT2) cells generate postmitotic neurons (NT2N cells) upon retinoic acid (RA) treatment and are functionally integrated in the host tissue following grafting into the rodent and human brain, thus representing a promising source for neuronal replacement therapy. Yet the major limitations of this model are the lengthy differentiation procedure and its low efficiency, although recent studies suggest that the differentiation process can be shortened to less than 1week using nucleoside analogues. To explore whether short-term exposure of NT2 cells to the nucleoside analogue cytosine β-d-arabinofuranoside (AraC) could be a suitable method to efficiently generate mature neurons, we conducted a neurochemical and morphometric characterization of AraC-differentiated NT2N (AraC/NT2N) neurons and improved the differentiation efficiency by modifying the cell culture schedule. Moreover, we analyzed the neurotransmitter phenotypes of AraC/NT2N neurons. Cultures obtained by treatment with AraC were highly enriched in postmitotic neurons and essentially composed of dual glutamatergic/cholinergic neurons, which contrasts with the preferential GABAergic phenotype that we found after RA differentiation.Taken together, our results further reinforce the notion NT2 cells are a versatile source of neuronal phenotypes and provide a new encouraging platform for studying mechanisms of neuronal differentiation and for exploring neuronal replacement strategies. |
| Databáze: | OpenAIRE |
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