Physiological Normoxia and Absence of EGF Is Required for the Long-Term Propagation of Anterior Neural Precursors from Human Pluripotent Cells

Bilada Bilican; Matthew R. Livesey; Ghazal Haghi; Jing Qiu; Karen Burr; Rick Siller; Giles E. Hardingham; David J. A. Wyllie; Siddharthan Chandran

doi:10.1371/journal.pone.0085932

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Research Article

Physiological Normoxia and Absence of EGF Is Required for the Long-Term Propagation of Anterior Neural Precursors from Human Pluripotent Cells

Bilada Bilican ,

Contributed equally to this work with: Bilada Bilican, Matthew R. Livesey

Affiliations Euan MacDonald Centre for Motor Neurone Disease Research, University of Edinburgh, Edinburgh, United Kingdom, MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom, Centre for Neuroregeneration, University of Edinburgh, Edinburgh, United Kingdom
⨯
Matthew R. Livesey ,

Contributed equally to this work with: Bilada Bilican, Matthew R. Livesey

Affiliations Euan MacDonald Centre for Motor Neurone Disease Research, University of Edinburgh, Edinburgh, United Kingdom, Centre for Neuroregeneration, University of Edinburgh, Edinburgh, United Kingdom, Centre for Integrative Physiology, University of Edinburgh, Edinburgh, United Kingdom
⨯
Ghazal Haghi,

Affiliations Euan MacDonald Centre for Motor Neurone Disease Research, University of Edinburgh, Edinburgh, United Kingdom, Centre for Neuroregeneration, University of Edinburgh, Edinburgh, United Kingdom, Centre for Integrative Physiology, University of Edinburgh, Edinburgh, United Kingdom
⨯
Jing Qiu,

Affiliation Centre for Integrative Physiology, University of Edinburgh, Edinburgh, United Kingdom
⨯
Karen Burr,

Affiliations Euan MacDonald Centre for Motor Neurone Disease Research, University of Edinburgh, Edinburgh, United Kingdom, MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom, Centre for Neuroregeneration, University of Edinburgh, Edinburgh, United Kingdom
⨯
Rick Siller,

Affiliations MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom, Centre for Neuroregeneration, University of Edinburgh, Edinburgh, United Kingdom
⨯
Giles E. Hardingham,

Affiliation Centre for Integrative Physiology, University of Edinburgh, Edinburgh, United Kingdom
⨯
David J. A. Wyllie ,

* E-mail: dwyllie1@staffmail.ed.ac.uk (DJAW); siddharthan.chandran@ed.ac.uk (SC)

Affiliation Centre for Integrative Physiology, University of Edinburgh, Edinburgh, United Kingdom
⨯
Siddharthan Chandran

* E-mail: dwyllie1@staffmail.ed.ac.uk (DJAW); siddharthan.chandran@ed.ac.uk (SC)

Affiliations Euan MacDonald Centre for Motor Neurone Disease Research, University of Edinburgh, Edinburgh, United Kingdom, MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom, Centre for Neuroregeneration, University of Edinburgh, Edinburgh, United Kingdom
⨯

Physiological Normoxia and Absence of EGF Is Required for the Long-Term Propagation of Anterior Neural Precursors from Human Pluripotent Cells

Bilada Bilican,
Matthew R. Livesey,
Ghazal Haghi,
Jing Qiu,
Karen Burr,
Rick Siller,
Giles E. Hardingham,
David J. A. Wyllie,
Siddharthan Chandran

Published: January 17, 2014
https://doi.org/10.1371/journal.pone.0085932

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Culture of neurons?

Posted by jasonblanton on 13 Mar 2014 at 11:19 GMT

The methods don't seem to describe the conditions under which the derived neurons were maintained. Were they kept at 3% O2? If not, how and when did the transition from 3% to ~21% occur? What CO2% was used?

thanks...
JB

No competing interests declared.

RE: Culture of neurons?

Bilada replied to jasonblanton on 13 Mar 2014 at 14:50 GMT

Thanks for your question.

The neurons were maintained at the respective oxygen concentrations that the NPCs were propagated and differentiated. Neurons were not transitioned from 3% to 21% O2, or vice a versa, after they were derived.

CO2% in both humidified tissue culture incubators (3% O2 and 21% O2) were set at 5%.

Regards,

Bilada

No competing interests declared.

RE: RE: Culture of neurons?

jasonblanton replied to Bilada on 15 Mar 2014 at 16:19 GMT

Excellent, thanks. Nicely done, BTW...

jb

No competing interests declared.

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Physiological Normoxia and Absence of EGF Is Required for the Long-Term Propagation of Anterior Neural Precursors from Human Pluripotent Cells

Physiological Normoxia and Absence of EGF Is Required for the Long-Term Propagation of Anterior Neural Precursors from Human Pluripotent Cells

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Culture of neurons?

Posted by jasonblanton on 13 Mar 2014 at 11:19 GMT

RE: Culture of neurons?

Bilada replied to jasonblanton on 13 Mar 2014 at 14:50 GMT

RE: RE: Culture of neurons?

jasonblanton replied to Bilada on 15 Mar 2014 at 16:19 GMT