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Predicting stem cell fate changes by differential cell cycle progression patterns
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| Zeitschriftentitel: | Development |
|---|---|
| Personen und Körperschaften: | , , , , , |
| In: | Development, 140, 2013, 2, S. 459-470 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
The Company of Biologists
|
| Schlagwörter: |
| author_facet |
Roccio, Marta Schmitter, Daniel Knobloch, Marlen Okawa, Yuya Sage, Daniel Lutolf, Matthias P. Roccio, Marta Schmitter, Daniel Knobloch, Marlen Okawa, Yuya Sage, Daniel Lutolf, Matthias P. |
|---|---|
| author |
Roccio, Marta Schmitter, Daniel Knobloch, Marlen Okawa, Yuya Sage, Daniel Lutolf, Matthias P. |
| spellingShingle |
Roccio, Marta Schmitter, Daniel Knobloch, Marlen Okawa, Yuya Sage, Daniel Lutolf, Matthias P. Development Predicting stem cell fate changes by differential cell cycle progression patterns Developmental Biology Molecular Biology |
| author_sort |
roccio, marta |
| spelling |
Roccio, Marta Schmitter, Daniel Knobloch, Marlen Okawa, Yuya Sage, Daniel Lutolf, Matthias P. 1477-9129 0950-1991 The Company of Biologists Developmental Biology Molecular Biology http://dx.doi.org/10.1242/dev.086215 <jats:p>Stem cell self-renewal, commitment and reprogramming rely on a poorly understood coordination of cell cycle progression and execution of cell fate choices. Using existing experimental paradigms, it has not been possible to probe this relationship systematically in live stem cells in vitro or in vivo. Alterations in stem cell cycle kinetics probably occur long before changes in phenotypic markers are apparent and could be used as predictive parameters to reveal changes in stem cell fate. To explore this intriguing concept, we developed a single-cell tracking approach that enables automatic detection of cell cycle phases in live (stem) cells expressing fluorescent ubiquitylation-based cell-cycle indicator (FUCCI) probes. Using this tool, we have identified distinctive changes in lengths and fluorescence intensities of G1 (red fluorescence) and S/G2-M (green) that are associated with self-renewal and differentiation of single murine neural stem/progenitor cells (NSCs) and embryonic stem cells (ESCs). We further exploited these distinctive features using fluorescence-activated cell sorting to select for desired stem cell fates in two challenging cell culture settings. First, as G1 length was found to nearly double during NSC differentiation, resulting in progressively increasing red fluorescence intensity, we successfully purified stem cells from heterogeneous cell populations by their lower fluorescence. Second, as ESCs are almost exclusively marked by the green (S/G2-M) FUCCI probe due to their very short G1, we substantially augmented the proportion of reprogramming cells by sorting green cells early on during reprogramming from a NSC to an induced pluripotent stem cell state. Taken together, our studies begin to shed light on the crucial relationship between cell cycle progression and fate choice, and we are convinced that the presented approach can be exploited to predict and manipulate cell fate in a wealth of other mammalian cell systems.</jats:p> Predicting stem cell fate changes by differential cell cycle progression patterns Development |
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Predicting stem cell fate changes by differential cell cycle progression patterns |
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Predicting stem cell fate changes by differential cell cycle progression patterns |
| title_full |
Predicting stem cell fate changes by differential cell cycle progression patterns |
| title_fullStr |
Predicting stem cell fate changes by differential cell cycle progression patterns |
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Predicting stem cell fate changes by differential cell cycle progression patterns |
| title_short |
Predicting stem cell fate changes by differential cell cycle progression patterns |
| title_sort |
predicting stem cell fate changes by differential cell cycle progression patterns |
| topic |
Developmental Biology Molecular Biology |
| url |
http://dx.doi.org/10.1242/dev.086215 |
| publishDate |
2013 |
| physical |
459-470 |
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<jats:p>Stem cell self-renewal, commitment and reprogramming rely on a poorly understood coordination of cell cycle progression and execution of cell fate choices. Using existing experimental paradigms, it has not been possible to probe this relationship systematically in live stem cells in vitro or in vivo. Alterations in stem cell cycle kinetics probably occur long before changes in phenotypic markers are apparent and could be used as predictive parameters to reveal changes in stem cell fate. To explore this intriguing concept, we developed a single-cell tracking approach that enables automatic detection of cell cycle phases in live (stem) cells expressing fluorescent ubiquitylation-based cell-cycle indicator (FUCCI) probes. Using this tool, we have identified distinctive changes in lengths and fluorescence intensities of G1 (red fluorescence) and S/G2-M (green) that are associated with self-renewal and differentiation of single murine neural stem/progenitor cells (NSCs) and embryonic stem cells (ESCs). We further exploited these distinctive features using fluorescence-activated cell sorting to select for desired stem cell fates in two challenging cell culture settings. First, as G1 length was found to nearly double during NSC differentiation, resulting in progressively increasing red fluorescence intensity, we successfully purified stem cells from heterogeneous cell populations by their lower fluorescence. Second, as ESCs are almost exclusively marked by the green (S/G2-M) FUCCI probe due to their very short G1, we substantially augmented the proportion of reprogramming cells by sorting green cells early on during reprogramming from a NSC to an induced pluripotent stem cell state. Taken together, our studies begin to shed light on the crucial relationship between cell cycle progression and fate choice, and we are convinced that the presented approach can be exploited to predict and manipulate cell fate in a wealth of other mammalian cell systems.</jats:p> |
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| author | Roccio, Marta, Schmitter, Daniel, Knobloch, Marlen, Okawa, Yuya, Sage, Daniel, Lutolf, Matthias P. |
| author_facet | Roccio, Marta, Schmitter, Daniel, Knobloch, Marlen, Okawa, Yuya, Sage, Daniel, Lutolf, Matthias P., Roccio, Marta, Schmitter, Daniel, Knobloch, Marlen, Okawa, Yuya, Sage, Daniel, Lutolf, Matthias P. |
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| description | <jats:p>Stem cell self-renewal, commitment and reprogramming rely on a poorly understood coordination of cell cycle progression and execution of cell fate choices. Using existing experimental paradigms, it has not been possible to probe this relationship systematically in live stem cells in vitro or in vivo. Alterations in stem cell cycle kinetics probably occur long before changes in phenotypic markers are apparent and could be used as predictive parameters to reveal changes in stem cell fate. To explore this intriguing concept, we developed a single-cell tracking approach that enables automatic detection of cell cycle phases in live (stem) cells expressing fluorescent ubiquitylation-based cell-cycle indicator (FUCCI) probes. Using this tool, we have identified distinctive changes in lengths and fluorescence intensities of G1 (red fluorescence) and S/G2-M (green) that are associated with self-renewal and differentiation of single murine neural stem/progenitor cells (NSCs) and embryonic stem cells (ESCs). We further exploited these distinctive features using fluorescence-activated cell sorting to select for desired stem cell fates in two challenging cell culture settings. First, as G1 length was found to nearly double during NSC differentiation, resulting in progressively increasing red fluorescence intensity, we successfully purified stem cells from heterogeneous cell populations by their lower fluorescence. Second, as ESCs are almost exclusively marked by the green (S/G2-M) FUCCI probe due to their very short G1, we substantially augmented the proportion of reprogramming cells by sorting green cells early on during reprogramming from a NSC to an induced pluripotent stem cell state. Taken together, our studies begin to shed light on the crucial relationship between cell cycle progression and fate choice, and we are convinced that the presented approach can be exploited to predict and manipulate cell fate in a wealth of other mammalian cell systems.</jats:p> |
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| spelling | Roccio, Marta Schmitter, Daniel Knobloch, Marlen Okawa, Yuya Sage, Daniel Lutolf, Matthias P. 1477-9129 0950-1991 The Company of Biologists Developmental Biology Molecular Biology http://dx.doi.org/10.1242/dev.086215 <jats:p>Stem cell self-renewal, commitment and reprogramming rely on a poorly understood coordination of cell cycle progression and execution of cell fate choices. Using existing experimental paradigms, it has not been possible to probe this relationship systematically in live stem cells in vitro or in vivo. Alterations in stem cell cycle kinetics probably occur long before changes in phenotypic markers are apparent and could be used as predictive parameters to reveal changes in stem cell fate. To explore this intriguing concept, we developed a single-cell tracking approach that enables automatic detection of cell cycle phases in live (stem) cells expressing fluorescent ubiquitylation-based cell-cycle indicator (FUCCI) probes. Using this tool, we have identified distinctive changes in lengths and fluorescence intensities of G1 (red fluorescence) and S/G2-M (green) that are associated with self-renewal and differentiation of single murine neural stem/progenitor cells (NSCs) and embryonic stem cells (ESCs). We further exploited these distinctive features using fluorescence-activated cell sorting to select for desired stem cell fates in two challenging cell culture settings. First, as G1 length was found to nearly double during NSC differentiation, resulting in progressively increasing red fluorescence intensity, we successfully purified stem cells from heterogeneous cell populations by their lower fluorescence. Second, as ESCs are almost exclusively marked by the green (S/G2-M) FUCCI probe due to their very short G1, we substantially augmented the proportion of reprogramming cells by sorting green cells early on during reprogramming from a NSC to an induced pluripotent stem cell state. Taken together, our studies begin to shed light on the crucial relationship between cell cycle progression and fate choice, and we are convinced that the presented approach can be exploited to predict and manipulate cell fate in a wealth of other mammalian cell systems.</jats:p> Predicting stem cell fate changes by differential cell cycle progression patterns Development |
| spellingShingle | Roccio, Marta, Schmitter, Daniel, Knobloch, Marlen, Okawa, Yuya, Sage, Daniel, Lutolf, Matthias P., Development, Predicting stem cell fate changes by differential cell cycle progression patterns, Developmental Biology, Molecular Biology |
| title | Predicting stem cell fate changes by differential cell cycle progression patterns |
| title_full | Predicting stem cell fate changes by differential cell cycle progression patterns |
| title_fullStr | Predicting stem cell fate changes by differential cell cycle progression patterns |
| title_full_unstemmed | Predicting stem cell fate changes by differential cell cycle progression patterns |
| title_short | Predicting stem cell fate changes by differential cell cycle progression patterns |
| title_sort | predicting stem cell fate changes by differential cell cycle progression patterns |
| title_unstemmed | Predicting stem cell fate changes by differential cell cycle progression patterns |
| topic | Developmental Biology, Molecular Biology |
| url | http://dx.doi.org/10.1242/dev.086215 |