Review Reports

Round 1

Reviewer 1 Report

This review by Dr. Jordi Soriano is a beautiful look at the past 15 years of great work coming from his lab. It is clearly written and well-presented, and the biophysical, computational, and mathematical concepts described are easy to follow. The review offers multiple examples of concepts and experiments that make it suitable for a broad audience in both research and medical communities. Finally, it not only provides up-to-date research venues but also future perspectives in this captivating field of neuroscience.

Minor typo: in Figure 1g, “cell therapy” instead of “cel therapy”.

Author Response

I appreciate the interest of the Reviewer in my work. I carried out the requested correction and revised the English language.

Reviewer 2 Report

The manuscript by Jordi Soriano titled “Neuronal cultures: exploring biophysics, complex systems, and medicine in a dish”. Is a systematic review in the field and gives a brief understanding of neuronal culture in medicine, can be considered for publication with minor edits. However, they need to address following comments.

 Major comments

 1.    Authors need to discuss the drawbacks/limitations/challenges of in-vitro neuronal cultures specifically hiPSCs in separate subheading  

2.    Subheading Brain on chip and future direction can be two different heading.

 

Minor comments:

 

1.    line 89 consider including maturity/aging as challenges in hiPSC culture in disease modeling

2.    Author had put so much effort in reviewing multiple aspects of neuronal cultures, but the resolution of image is not clear. Hence, I request author to increase the resolution or size of each figure other than figure 3.

3.    Line 17- Weather- Whether

4.    line 4 neuroophysics- neurophysics

5.    line 81 Cells lines’- cell lines

6.    Please relevant add the fluorescent dyes used in the experiments in figure legends

7.    If possible, add in short Electrophysiological / other instrument details in the figure legends  

8.    61^st reference need to be checked

9.    12,13, 20, 24,25,27,41,45,53,64,81,82,83—DOI need to be included

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Reviewer 3 Report

This review summarize the author's work and main achievements in the last 15 years. The paper is complexively well presented: it starts with the neuronal culture (primary rodent and human iPSCs-derived) overview and it continues with multidisciplinary approach that spread in several lines of research, allowed numerous publications and opened promissing applications in biomedicine. Historical overview, experimental and theoretical parts are clearly described and the review could be (relatively easily) read and understood by non-experts (for instance, section 5 is well explained so that researchers with biomedical background without computational skills can capture the main points). 

There are just few points that require major attention:

Line 64-65: The word deceased can be removed since by saying „neurons are extracted“ it is already clear that the animals are sacrificed before the tissue extraction. The statement „before any deterioration of tissue occurs” is not true – the deterioration of the tissue starts immediately following euthananasia and in particular during the mechanical and enzymatic tissue dissociation. Even if one tries to extract the tissue as quickly as possible, the dissection requires several minutes during which the neurons start to die. (The timing of all steps in the most frequently using protocols is reported in Beaudoin et al, 2012 Nat Protoc. 2012 Sep;7(9):1741-54. doi: 10.1038/nprot.2012.099). It would be more correct to state that the animal tissue (brain) is freshly isolated to avoid (or minimize) as much as possible tissue deterioration.

Line 68: The sentence „Neurons are seeded on the substrate together with glia” should be corrected since when embryonic rodent tissue is used to prepare neuronal culture (for example, in the ref. no. 12, E18-19 rat cortex was used), glial cells are still not developed (gliogenesis starts postnatally in rodents). In these cases, nearly pure neuronal cultures are formed and glial cells develop only later in the following days/weeks in vitro (in the presence of growth factors contained in serum). 

Line 96: In the figure legend of the Figure 2a (right panel) and c, the description of the labeling method (fluorescent calcium dye?) used for fluorescence image is missing. This is reported later in the main text (Line 106 page 3). Figure 2b: Electrodes are visible as black circles and lines.

Minor points:

Line 4: typing error (neuroophysics)

Line 25: conditions? (instead of environment) – to avoid repeating of the same word

Line 36: perhaps alliance is more neutral word, retaining the same meaning as marriage

Line 63: not completely true that after few days in vitro the spontaneous activity is abundant (the first week in vitro usually presents first signs of relatively weak activity, but in the following two weeks significantly higher activity is usually observed (in terms of spikes amplitude and frequency) due to the network maturation)

Line 86: I suggest to cite the original works (or at least the one involving human cell reprogramming):

Takahashi, K.; Yamanaka, S. Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors. Cell 2006, 126, 663–676, doi:10.1016/j.cell.2006.07.024.

Takahashi, K.; Tanabe, K.; Ohnuki, M.; Narita, M.; Ichisaka, T.; Tomoda, K.; Yamanaka, S. Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors. Cell 2007, 131, 861–872, doi:10.1016/j.cell.2007.11.019.

Line 94: „to get data fast with the latter” rephrasing required?

Line 123: check the Journal's guidelines – is it necessary to mention/cite the institute? (usually the author's first name or only reference number is cited in the main text)

Line 155: The sentence „We also considered in general networks relatively large….” requires rephrasing. 

Line 452: controlled environment (instead of „control“)?

 

Author Response

Please see the attachment.

Author Response File: Author Response.pdf