A Neurofilament-L reporter cell line for the quantification of early neuronal differentiation: A Bioassay for neurotrophic activities
Background: Neurotrophic activity constitutes a crucial factor in the recovery from neurological injuries and is impaired in neurodegenerative disorders. Preclinical studies of neurotrophic factors to improve outcome of neurodegenerative diseases have yielded promising results. However, due to the c...
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| Format: | Article |
| Language: | English |
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Elsevier
2024-01-01
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| Series: | Heliyon |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844024007849 |
| _version_ | 1797328634580566016 |
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| author | Lisa-Franziska Seidl Stefan Winter Ludwig Aigner Julia Schartner |
| author_facet | Lisa-Franziska Seidl Stefan Winter Ludwig Aigner Julia Schartner |
| author_sort | Lisa-Franziska Seidl |
| collection | DOAJ |
| description | Background: Neurotrophic activity constitutes a crucial factor in the recovery from neurological injuries and is impaired in neurodegenerative disorders. Preclinical studies of neurotrophic factors to improve outcome of neurodegenerative diseases have yielded promising results. However, due to the complexity of these therapies, the clinical translation of this approach was so far not successful and more feasible treatments with neurotrophic activity may be promising alternatives. Therefore, highly sensitive and robust assays for compound screening are required. New method: Nerve growth factor is known to induce Neurofilament-L (NF-L) expression in a rat pheochromocytoma cell line (PC12 cells) during early neuronal differentiation. We generated and characterized an enhanced green fluorescent protein (EGFP)-NF-L reporter PC12 cell line for the development of a cell-based assay (designated Neurofilament-L Bioassay) that allows straightforward quantification of early neuronal differentiation based on NF-L expression. Results: Using Cerebrolysin® as a role model for a pharmacological compound that stimulates neurotrophic activity in the central nervous system, the Neurofilament-L Bioassay was proved to be a robust, specific, and reproducible method. Comparison with existing method(s): It was already shown that NF-L expression correlates with neurite outgrowth in PC12 cells. Currently, quantification of neurite outgrowth is the most commonly used method to evaluate neuronal differentiation in PC12 cells, an approach that is time-consuming and of high variability. Conclusions: This work describes the development of an EGFP-NF-L reporter PC12 cell-based assay as a robust and reproducible tool for “high throughput” compound screening for neurotrophic activity. |
| first_indexed | 2024-03-08T06:54:31Z |
| format | Article |
| id | doaj.art-faf4204e901a4c74a586c5c87e7117ad |
| institution | Directory Open Access Journal |
| issn | 2405-8440 |
| language | English |
| last_indexed | 2024-03-08T06:54:31Z |
| publishDate | 2024-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj.art-faf4204e901a4c74a586c5c87e7117ad2024-02-03T06:38:22ZengElsevierHeliyon2405-84402024-01-01102e24753A Neurofilament-L reporter cell line for the quantification of early neuronal differentiation: A Bioassay for neurotrophic activitiesLisa-Franziska Seidl0Stefan Winter1Ludwig Aigner2Julia Schartner3Institute of Molecular Regenerative Medicine, Paracelsus Medical University, Salzburg, Austria; EVER Neuro Pharma, Oberburgau 3, 4866 Unterach, AustriaEVER Neuro Pharma, Oberburgau 3, 4866 Unterach, AustriaInstitute of Molecular Regenerative Medicine, Paracelsus Medical University, Salzburg, AustriaEVER Neuro Pharma, Oberburgau 3, 4866 Unterach, Austria; Corresponding author.Background: Neurotrophic activity constitutes a crucial factor in the recovery from neurological injuries and is impaired in neurodegenerative disorders. Preclinical studies of neurotrophic factors to improve outcome of neurodegenerative diseases have yielded promising results. However, due to the complexity of these therapies, the clinical translation of this approach was so far not successful and more feasible treatments with neurotrophic activity may be promising alternatives. Therefore, highly sensitive and robust assays for compound screening are required. New method: Nerve growth factor is known to induce Neurofilament-L (NF-L) expression in a rat pheochromocytoma cell line (PC12 cells) during early neuronal differentiation. We generated and characterized an enhanced green fluorescent protein (EGFP)-NF-L reporter PC12 cell line for the development of a cell-based assay (designated Neurofilament-L Bioassay) that allows straightforward quantification of early neuronal differentiation based on NF-L expression. Results: Using Cerebrolysin® as a role model for a pharmacological compound that stimulates neurotrophic activity in the central nervous system, the Neurofilament-L Bioassay was proved to be a robust, specific, and reproducible method. Comparison with existing method(s): It was already shown that NF-L expression correlates with neurite outgrowth in PC12 cells. Currently, quantification of neurite outgrowth is the most commonly used method to evaluate neuronal differentiation in PC12 cells, an approach that is time-consuming and of high variability. Conclusions: This work describes the development of an EGFP-NF-L reporter PC12 cell-based assay as a robust and reproducible tool for “high throughput” compound screening for neurotrophic activity.http://www.sciencedirect.com/science/article/pii/S2405844024007849BioassayCerebrolysin®Nerve growth factorNeuronal differentiationNeuroplasticityNeurotrophins |
| spellingShingle | Lisa-Franziska Seidl Stefan Winter Ludwig Aigner Julia Schartner A Neurofilament-L reporter cell line for the quantification of early neuronal differentiation: A Bioassay for neurotrophic activities Heliyon Bioassay Cerebrolysin® Nerve growth factor Neuronal differentiation Neuroplasticity Neurotrophins |
| title | A Neurofilament-L reporter cell line for the quantification of early neuronal differentiation: A Bioassay for neurotrophic activities |
| title_full | A Neurofilament-L reporter cell line for the quantification of early neuronal differentiation: A Bioassay for neurotrophic activities |
| title_fullStr | A Neurofilament-L reporter cell line for the quantification of early neuronal differentiation: A Bioassay for neurotrophic activities |
| title_full_unstemmed | A Neurofilament-L reporter cell line for the quantification of early neuronal differentiation: A Bioassay for neurotrophic activities |
| title_short | A Neurofilament-L reporter cell line for the quantification of early neuronal differentiation: A Bioassay for neurotrophic activities |
| title_sort | neurofilament l reporter cell line for the quantification of early neuronal differentiation a bioassay for neurotrophic activities |
| topic | Bioassay Cerebrolysin® Nerve growth factor Neuronal differentiation Neuroplasticity Neurotrophins |
| url | http://www.sciencedirect.com/science/article/pii/S2405844024007849 |
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