Quantification of Muscle Satellite Stem Cell Divisions by High-Content Analysis
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature..
High-content screening is commonly performed on 2D cultured cells, which is high throughput but has low biological relevance. In contrast, single myofiber culture assay preserves the satellite cell niche between the basal lamina and sarcolemma and consequently has high biological relevance but is low throughput. We describe here a high-content screening method that utilizes single myofiber culture that addresses the caveats of both techniques. Our method utilizes the transgenic reporter allele Myf5-Cre:R26R-eYFP to differentiate stem and committed cells within a dividing couplet that can be quantified by high-content throughput immunodetection and bioinformatic analysis.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:2587 |
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| Enthalten in: |
Methods in molecular biology (Clifton, N.J.) - 2587(2023) vom: 18., Seite 537-553 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Chen, William [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 22.11.2022 Date Revised 29.11.2022 published: Print Citation Status MEDLINE |
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| doi: |
10.1007/978-1-0716-2772-3_29 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 520 | |a High-content screening is commonly performed on 2D cultured cells, which is high throughput but has low biological relevance. In contrast, single myofiber culture assay preserves the satellite cell niche between the basal lamina and sarcolemma and consequently has high biological relevance but is low throughput. We describe here a high-content screening method that utilizes single myofiber culture that addresses the caveats of both techniques. Our method utilizes the transgenic reporter allele Myf5-Cre:R26R-eYFP to differentiate stem and committed cells within a dividing couplet that can be quantified by high-content throughput immunodetection and bioinformatic analysis | ||
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| 650 | 4 | |a Symmetric division | |
| 700 | 1 | |a Perkins, Theodore J |e verfasserin |4 aut | |
| 700 | 1 | |a Rudnicki, Michael A |e verfasserin |4 aut | |
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