Proline-based solution maintains cell viability and stemness of canine adipose-derived mesenchymal stem cells after hypothermic storage

Pongsatorn Horcharoensuk; Sunantha Yang-en; Amarin Narkwichean; Ruttachuk Rungsiwiwut

doi:10.1371/journal.pone.0264773

Peer Review History

Original SubmissionOctober 5, 2021
15 Dec 2021 Decision Letter - Nazmul Haque, Editor PONE-D-21-31968Proline-based solution maintains cell viability and stemness of canine adipose-derived mesenchymal stem cells after hypothermic storagePLOS ONE Dear Dr. Rungsiwiwut, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Please submit your revised manuscript by Jan 29 2022 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. 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Any potentially identifying patient information must be fully anonymized. Important: If there are ethical or legal restrictions to sharing your data publicly, please explain these restrictions in detail. Please see our guidelines for more information on what we consider unacceptable restrictions to publicly sharing data: http://journals.plos.org/plosone/s/data-availability#loc-unacceptable-data-access-restrictions. Note that it is not acceptable for the authors to be the sole named individuals responsible for ensuring data access. We will update your Data Availability statement to reflect the information you provide in your cover letter. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: No ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: No ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: The manuscript (PONE-D-21-31968_reviewer) The manuscript demonstrates best conditions to store mesenchymal stem cells (MSCs) meant for clinical or translation application. The paper highlights proline ability to protect MSCs from apoptosis within 6hrs of hypothermic conditions. The proline protection extend to improve proliferation, and stemness ability of MSCs. The conclusion was that proline will be beneficial in case of short term transportation of MSCs. Problems with the manuscript. • Typos and errors are not acceptable such as “hu-man”, “col-leagues”, “sur-face”, “Live-” and others. Authors could benefit from professional editing services. • For mitochondrial membrane potential assay authors should mention how many cells were loaded in 96 well-plate. • For proliferation assay, authors should mention how many cells were loaded in 96 well-plate. • Adding the figure legends between texts is confusing. Authors should add the legends at the end of manuscript after references. • Authors should mention how many cells have been gated during flow cytometry experiments. • Fig. 4 needs a scale bar to be added. • Logical error when authors wrote, “when cAD-MSCs are stored in NSS and 1 mM PL-BS” It should be “when cAD-MSCs are stored in NSS or 1 mM PL-BS” since they are not combined together. Authors could use professional editing services. • The authors claim that proline protected the mitochondria dysfunction. The present study did not support this notion. The sentence should be removed or modified into less affirm tone. • The qPCR raw data need to be available as supplementary such as Fig.5. • In line 494, authors should add “maybe” before “by preserving mitochondrial function”. Minor issues • Minor English punctuation errors. I recommend major revision to the current version ****** 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: Yes:** Bahauddeen Alrfaei [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0264773.r001
Revision 1
19 Jan 2022 Author Response Response to Editor and Journal requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf Answer: We provided the manuscript according to PLOS ONE style requirements. 2. We suggest you thoroughly copyedit your manuscript for language usage, spelling,and grammar. If you do not know anyone who can help you do this, you may wish to consider employing a professional scientific editing service. Whilst you may use any professional scientific editing service of your choice, PLOS has partnered with both American Journal Experts (AJE) and Editage to provide discounted services to PLOS authors. Both organizations have experience helping authors meet PLOS guidelines and can provide language editing, translation, manuscript formatting, and figure formatting to ensure your manuscript meets our submission guidelines. To take advantage of our partnership with AJE, visit the AJE website (http://aje.com/go/plos) for a 15% discount off AJE services. To take advantage of our partnership with Editage, visit the Editage website (www.editage.com) and enter referral code PLOSEDIT for a 15% discount off Editage services. If the PLOS editorial team finds any language issues in text that either AJE or Editage has edited, the service provider will re-edit the text for free. Upon resubmission, please provide the following: The name of the colleague or the details of the professional service that edited your manuscript. Answer: The language of our manuscript had been edited by the professional English-language editing staff at International College for Sustainability Studies of Srinakharinwirot University. The certificate of language editing is attached as a PDF. A copy of your manuscript showing your changes by either highlighting them or using track changes (uploaded as a supporting information file) Answer: Highlighting revised manuscript is provided and uploaded as a supporting information file. A clean copy of the edited manuscript (uploaded as the new manuscript file Answer: A clean edited manuscript is provided and uploaded as the new manuscript file. 3. Thank you for stating the following financial disclosure: “This research was funded by the Faculty of Medicine, Srinakharinwirot University (MED-GRAD-150; 511/2563 and MED-RES-200; 217/2563).” Please state what role the funders took in the study. If the funders had no role, please state: " The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." If this statement is not correct you must amend it as needed. Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf. Answer: Due to the funders had no role in the present study, therefore, we state “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript”. 4. Thank you for stating the following in the Acknowledgments Section of your manuscript: “This research was funded by the Faculty of Medicine, Srinakharinwirot University (MED-GRAD-150; 511/2563 and MED-RES-200; 217/2563).” We note that you have provided information within the Acknowledgements Section. Please note that funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: This research was funded by the Faculty of Medicine, Srinakharinwirot University (MED-GRAD-150; 511/2563 and MED-RES-200; 217/2563). Answer: The funding statement has been removed from the Acknowledgment section. Please include your amended statements within your cover letter; we will change the online submission form on your behalf. 5. Please upload a new copy of Figure 1A as the detail is not clear. Please follow the link for more information: https://blogs.plos.org/plos/2019/06/looking-good-tips-for-creating-your-plos-figures-graphics/ Answer: The resolution of Figure 1A has been improved. 6. In your Data Availability statement, you have not specified where the minimal data set underlying the results described in your manuscript can be found. PLOS defines a study's minimal data set as the underlying data used to reach the conclusions drawn in the manuscript and any additional data required to replicate the reported study findings in their entirety. All PLOS journals require that the minimal data set be made fully available. Answer: We uploaded the data set as the supporting information file (S2) For more information about our data policy, please see http://journals.plos.org/plosone/s/data-availability. Upon re-submitting your revised manuscript, please upload your studys minimal underlying data set as either Supporting Information files or to a stable, public repository and include the relevant URLs, DOIs, or accession numbers within your revised cover letter. For a list of acceptable repositories, please see http://journals.plos.org/plosone/s/data-availability#loc-recommended-repositories. Any potentially identifying patient information must be fully anonymized. Important: If there are ethical or legal restrictions to sharing your data publicly, please explain these restrictions in detail. Please see our guidelines for more information on what we consider unacceptable restrictions to publicly sharing data: http://journals.plos.org/plosone/s/data-availability#loc-unacceptable-data-access-restrictions. Note that it is not acceptable for the authors to be the sole named individuals responsible for ensuring data access. We will update your Data Availability statement to reflect the information you provide in your cover letter. Reviewer #1: The manuscript (PONE-D-21-31968_reviewer) The manuscript demonstrates best conditions to store mesenchymal stem cells (MSCs) meant for clinical or translation application. The paper highlights proline ability to protect MSCs from apoptosis within 6hrs of hypothermic conditions. The proline protection extends to improve proliferation, and stemness ability of MSCs. The conclusion was that proline will be beneficial in case of short-term transportation of MSCs. Problems with the manuscript. Typos and errors are not acceptable such as “hu-man”, “col-leagues”, “sur-face”, “Live-” and others. Authors could benefit from professional editing services. • Answer: All typos and errors have been corrected in the revised manuscript by professional grammar editing services from the International College for Sustainability Studies, Srinakharinwirot University. For mitochondrial membrane potential assay authors should mention how many cells were loaded in 96 well-plates. • Answer: We added the cell number (1x105 cells/well) that was used in the mitochondrial membrane potential assay as the reviewer suggested. The cell number appear in line 168 of the revised manuscript. For proliferation assay, authors should mention how many cells were loaded in 96 well-plates. • Answer: We added the cell number (1x104 cells/well) that was used in the proliferation assay as the reviewer suggested. The cell number appear in line 175 of the revised manuscript. Adding the figure legends between texts is confusing. Authors should add the legends at the end of manuscript after references. • Answer: We moved all of the figure legends to the end of the revised manuscript (lines 639–715) as the reviewer suggested. Authors should mention how many cells have been gated during flow cytometry experiments. • Answer: The number of cells that were used in the flow cytometry assay was added into both the cell viability and the apoptotic/necrotic fraction assay and the expression of specific surface antigens assay, which appeared in lines 163 and 191 of the revised manuscript. Fig. 4 needs a scale bar to be added. • Answer: The scale bars were embedded in Fig 4 as the reviewer suggested. Logical error when authors wrote, “when cAD-MSCs are stored in NSS and 1 mM PL-BS” It should be “when cAD-MSCs are stored in NSS or 1 mM PL-BS” since they are not combined together. Authors could use professional editing services. • Answer: We amended the texts into “when cAD-MSCs are stored in NSS or 1 mM PL-BS” which is shown in line 675 of the revised manuscript. The authors claim that proline protected the mitochondria dysfunction. The present study did not support this notion. The sentence should be removed or modified into less affirm tone. • Answer: We amended the sentence “proline protected the mitochondria dysfunction” to “proline might maintain mitochondrial integrity” which is shown in line 394-395 of the revised manuscript. The qPCR raw data need to be available as supplementary such as Fig.5. • Answer: We added the qPCR data in supplementation S2 Table as the reviewer suggested, which appears in lines 712-715. In line 494, authors should add “maybe” before “by preserving mitochondrial function”. • Answer: The word "maybe" was added to the sentence before "by preserving mitochondrial function" as the reviewer suggested, and it appears in line 427 of the revised manuscript. Minor issues Minor English punctuation errors. • Answer: The revised manuscript has undergone grammatical correction by professional grammar editing services from International College for Sustainability Studies, Srinakharinwirot University. Attachments Attachment Submitted filename: Response to the Journal requirements.docx https://doi.org/10.1371/journal.pone.0264773.r002
17 Feb 2022 Decision Letter - Nazmul Haque, Editor Proline-based solution maintains cell viability and stemness of canine adipose-derived mesenchymal stem cells after hypothermic storage PONE-D-21-31968R1 Dear Dr. Rungsiwiwut, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Nazmul Haque Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #1: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes ****** 6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: (No Response) ****** 7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: Yes:** Bahauddeen Alrfaei https://doi.org/10.1371/journal.pone.0264773.r003
Formally Accepted
21 Feb 2022 Acceptance Letter - Nazmul Haque, Editor PONE-D-21-31968R1 Proline-based solution maintains cell viability and stemness of canine adipose-derived mesenchymal stem cells after hypothermic storage Dear Dr. Rungsiwiwut: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Nazmul Haque Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0264773.r004

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