Physiological normoxia and absence of EGF is required for the long-term propagation of anterior neural precursors from human pluripotent cells.
Widespread use of human pluripotent stem cells (hPSCs) to study neuronal physiology and function is hindered by the ongoing need for specialist expertise in converting hPSCs to neural precursor cells (NPCs). Here, we describe a new methodology to generate cryo-preservable hPSC-derived NPCs that reta...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0085932 |
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| author | Bilada Bilican Matthew R Livesey Ghazal Haghi Jing Qiu Karen Burr Rick Siller Giles E Hardingham David J A Wyllie Siddharthan Chandran |
| author_facet | Bilada Bilican Matthew R Livesey Ghazal Haghi Jing Qiu Karen Burr Rick Siller Giles E Hardingham David J A Wyllie Siddharthan Chandran |
| author_sort | Bilada Bilican |
| collection | DOAJ |
| description | Widespread use of human pluripotent stem cells (hPSCs) to study neuronal physiology and function is hindered by the ongoing need for specialist expertise in converting hPSCs to neural precursor cells (NPCs). Here, we describe a new methodology to generate cryo-preservable hPSC-derived NPCs that retain an anterior identity and are propagatable long-term prior to terminal differentiation, thus abrogating regular de novo neuralization. Key to achieving passagable NPCs without loss of identity is the combination of both absence of EGF and propagation in physiological levels (3%) of O2. NPCs generated in this way display a stable long-term anterior forebrain identity and importantly retain developmental competence to patterning signals. Moreover, compared to NPCs maintained at ambient O2 (21%), they exhibit enhanced uniformity and speed of functional maturation, yielding both deep and upper layer cortical excitatory neurons. These neurons display multiple attributes including the capability to form functional synapses and undergo activity-dependent gene regulation. The platform described achieves long-term maintenance of anterior neural precursors that can give rise to forebrain neurones in abundance, enabling standardised functional studies of neural stem cell maintenance, lineage choice and neuronal functional maturation for neurodevelopmental research and disease-modelling. |
| format | Article |
| id | doaj-art-71432849ec4347e083d21fefa0fd69b9 |
| institution | DOAJ |
| issn | 1932-6203 |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-71432849ec4347e083d21fefa0fd69b92025-08-20T03:11:26ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0191e8593210.1371/journal.pone.0085932Physiological normoxia and absence of EGF is required for the long-term propagation of anterior neural precursors from human pluripotent cells.Bilada BilicanMatthew R LiveseyGhazal HaghiJing QiuKaren BurrRick SillerGiles E HardinghamDavid J A WyllieSiddharthan ChandranWidespread use of human pluripotent stem cells (hPSCs) to study neuronal physiology and function is hindered by the ongoing need for specialist expertise in converting hPSCs to neural precursor cells (NPCs). Here, we describe a new methodology to generate cryo-preservable hPSC-derived NPCs that retain an anterior identity and are propagatable long-term prior to terminal differentiation, thus abrogating regular de novo neuralization. Key to achieving passagable NPCs without loss of identity is the combination of both absence of EGF and propagation in physiological levels (3%) of O2. NPCs generated in this way display a stable long-term anterior forebrain identity and importantly retain developmental competence to patterning signals. Moreover, compared to NPCs maintained at ambient O2 (21%), they exhibit enhanced uniformity and speed of functional maturation, yielding both deep and upper layer cortical excitatory neurons. These neurons display multiple attributes including the capability to form functional synapses and undergo activity-dependent gene regulation. The platform described achieves long-term maintenance of anterior neural precursors that can give rise to forebrain neurones in abundance, enabling standardised functional studies of neural stem cell maintenance, lineage choice and neuronal functional maturation for neurodevelopmental research and disease-modelling.https://doi.org/10.1371/journal.pone.0085932 |
| spellingShingle | Bilada Bilican Matthew R Livesey Ghazal Haghi Jing Qiu Karen Burr Rick Siller Giles E Hardingham David J A Wyllie Siddharthan Chandran Physiological normoxia and absence of EGF is required for the long-term propagation of anterior neural precursors from human pluripotent cells. PLoS ONE |
| title | Physiological normoxia and absence of EGF is required for the long-term propagation of anterior neural precursors from human pluripotent cells. |
| title_full | Physiological normoxia and absence of EGF is required for the long-term propagation of anterior neural precursors from human pluripotent cells. |
| title_fullStr | Physiological normoxia and absence of EGF is required for the long-term propagation of anterior neural precursors from human pluripotent cells. |
| title_full_unstemmed | Physiological normoxia and absence of EGF is required for the long-term propagation of anterior neural precursors from human pluripotent cells. |
| title_short | Physiological normoxia and absence of EGF is required for the long-term propagation of anterior neural precursors from human pluripotent cells. |
| title_sort | physiological normoxia and absence of egf is required for the long term propagation of anterior neural precursors from human pluripotent cells |
| url | https://doi.org/10.1371/journal.pone.0085932 |
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