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Based on the reviews provided by two experts we found the article to potentially benefit the scientific community. Looking forward to its publication.
This is an excellent study that compares the human neural progenitor cells grown in physioxia and normoxia in terms of growth kinetics, glycolytic metabolism, mitochondrial content, mitochondrial membrane potential, oxygen uptake, ATP production, ROS production, and antioxidant enzymatic activity, thus shedding light on the importance of oxygenation control in studies focusing on disease and drug screening assays.
The authors have done a thoughtful experimental design and the revised article is clearly explained.
Authors have evidently demonstrated that alterations in physioxia-grown NPCs at the metabolic level not only occur but also lead to a differential response to exogenous oxidative agents.
Authors attempted to address the concerns raised and satisfactorily addressed the concerns.
Authors did a good job to address the concerns raised.
From the reviews it is evident that the manuscript has received potential scientific interest. The authors are suggested to address minor concerns raised by reviewers 1 and 3. Looking forward to your resubmission.
This study investigates the importance of oxygen levels in neuronal development which will be used for drug screening. The authors showed that altered morphology of human neural progenitor cells and oxidative functions in physioxia condition. They also demonstrate that changes in the mitochondrial physiology in human neural progenitor cells are induced by O2 availability. They showed that human neural progenitor cells cultured in 3% oxygen and exposed to external agents have greater susceptibility to DNA damage.
The authors did an extensive and a thoughtful in vitro analysis to demonstrate the role of oxygen levels in the development of human neural progenitor cells.
Overall, this is an interesting study. Interpretations of all the results by the authors are fully supported by the experimental data.
I would like to accept this manuscript for publication.
Experimental were well designed and performed.
This is an interesting and well planned study. Authors have shown that, oxygen levels should be carefully considered for in vitro models of neuronal development and drug screening.
i have few concerns
-Abstract should be reframed, as it is confusing at this moment.
-Authors should include the figures of the NPCs differentiation from human ESCs and their characterization in the manuscript. Which are lacking at this moment.
- Authors should see the effect of antimycin-C on cell death, as authors have shown elevated ROS levels in 21% oxygen (in Fig.5)
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