Entomoculture: A Preliminary Techno-Economic Assessment

However, I believe it is still needed to include more explanations on what the hypothetical product yielded by the insect cell cultivation would be, would require significant post-processing to get to the stage where it resembles traditional meat in both texture and taste.

Added clarification of products to methods section, line 117-118. See below. Added reference to this as a limitation in the conclusion, lines 461-464. “Further research into this topic may explore costs associated with scaffolding or downstream processing associated with achieving more conventional-meat-like products (i.e., 3-D structure, texture, flavoring, etc.).”

It is not mentioned anywhere if the authors refer to only minced meat-like product as the end result of the insect cell cultivation? What about 3D steak-like products? If they are considered as well, then the use of scaffolds and associated costs of biomaterials, fabrication methods and other types of bioreactors should be counted in. it should be described if any support structures are intended for use for insect-based cultivated meat

Added justification for the omission of scaffolds and microcarriers to the methods section, lines 115-120. “The cost model should be interpreted as a preliminary TEA, as certain processes such as seed train, scaffolding, microcarriers, and bioreactor cleaning are omitted. Downstream and post processing are also not considered within the original or present models, so the hypothetical product systems are assumed to yield products similar to minced meat. While these are certainly important factors for consideration in the cost estimation of cultured meat, maintaining included or excluded processes in the model constant between studies allows for comparison between the two cell types, which was the objective of the present study.”

Another point which I find somewhat surprising is that the Antheraea clone “AeC6" were included only in the section when discussing the cell dimensions. Seeing that their use appears to significantly bring the total costs down, one needs to ask why are they not included in the full analyses in the same way as Hi-Five/Sf9 or S2 cells?

Robust data on necessary parameters for the model were only available for Hi-Five, Sf9, and S2 cells.

Spelling: line 9 - nutritious instead of nutritous

line 95 - presents instead of present

This has been fixed.

it is not clear why the authors decided to use the open-source ACMB cost model developed by Risner et al.

This cost model was purposely made open-source for use by the public to compare cultured meat product systems. This is stated in lines 84-86.

why the choice of lepidopteran ovarian cells

Two lepidopteran cell lines and one Drosophila cell line were used due to data availability. Lines 124-129 were edited to convey the reasoning: “Three insect cell lines were the focus of this study— Sf-9, Hi-Five, and S2— chosen for their common use in the pharmaceutical industry and robust data availability. Both originate from lepidopteran ovarian cells, the Sf-9 and Hi-Five cell lines had similar characteristics and were therefore analyzed together. S2 cells, which were established from Drosophila melanogaster embryonic tissue and are the most used Drosophila cell line, were analyzed independently due to their unique characteristics.”

In order to read the results, the authors could codify (maybe in the same table) the costs of each component in different scenarios: bio-equipment and media, electricity, water, labor. 

Table 2 was added to directly compare costs of components. Lines 180-185 were added to more broadly discuss results before deeper analysis.

the conclusions are very similar to the results and there is no reflection concerning why this paper could be useful for industrial companies. It could be useful add some reflections about future research need linked to the topic and limitations of this study.

Section 5 has been edited to address this, and reflections on limitations and future directions have been added.

It should be stressed which factors mostly effect the costs (energy, raw materials)

Added lines 180-186 and more text about this in the conclusion. 

Results and discussion section the "understanding model outcomes" paragraphs 3.1 to 3.6 describe methods used to define models and to analyze data. These should be moved to materials and methods section.

Section 2.1 describes methods used to define insect models, sections 3.1-3.6 discuss the major outcome differences between scenarios while tracking and highlighting differences in the model to explain these differences

Paragraph number 1.3 was already used in the previous one.

This has been fixed.

2 cell lines?

Changed to two cell types (insect and mammalian)

Here you state that the production of cultivated meat should avoid serum of animal origins and in particular FBS. However previously (line 99) you reported that the possibility to grow in serum free media was an advantage for insect cell lines. Please explain and check the text for consistency about this point.

Lines 417-426 reworded for clarity.

“Oftentimes, cell cultures will be supplemented with fetal bovine serum (FBS), but this is a byproduct of the meat industry, while in contrast, this model assumes the use of recombinant growth factors as animal serum alternatives. Cell cultures for cultivated meat can instead utilize these animal-free growth factors to increase proliferation, but as seen in the original model, these impose some of the most significant costs to production. As emphasized earlier in the text, insect cells can grow in the absence of these costly growth factors included in the original mammalian cell-based model, such as transforming growth factor beta and fibroblast growth factor 2. As increased cell concentration was found to be a considerable cost lever, other growth factors specific to insect cells have been identified and may be added to promote growth while not significantly contributing to cost of production.”