Review Reports

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The proposed manuscript by Ralf Weiskirchen is a narrative review that documents the history, technical state-of-the-art, economic impact, regulatory environment, and practical recommendations for routine cell-line authentication (primarily STR profiling) and complementary approaches (SNP panels, low-pass WGS, dPCR, methylation clocks).

It frames authentication as both scientific best practice and an economic imperative for reproducibility.

 

According to the author, STR profiling (modern 24-plex kits + six-dye capillary electrophoresis) remains the practical gold standard for human cell-line authentication: fast, inexpensive and highly discriminatory when benchmarked against curated databases. The manuscript gives the technical rationale (marker sets, interpretation thresholds, QC considerations). Complementary methods (SNP panels, low-pass WGS, digital PCR, methylation/epigenetic clocks) fill blind spots (aneuploidy, mixed species, copy-number changes, “age”/epigenetic drift) and are inexpensive enough to be considered routine in many settings. The aggregate economic case is strong: irreproducible research (with cell-line misidentification as a major contributor) imposes very large costs (cited estimate: ~$28 billion/year in the U.S.), and simple ROI models in the review indicate routine STR testing is highly cost-effective at laboratory and institutional levels. The paper includes example cost-benefit tables and an STR profiling data prompt. Policy and publishing landscapes are shifting: ANSI/ATCC standards, NIH requirements for resource authentication in grants, journal policies (AACR, Nature family, others), and regulatory (FDA/EMA) guidance increasingly expect/require authentication. Practical recommendations and a roadmap are provided (e.g., gatekeeper rules, schedule suggestions, FAIR-compliant metadata, LIMS integration, periodic testing, combination of STR + mycoplasma + species checks).

The author concludes that routine, documented authentication (STR + targeted complementary assays) is now technically feasible, economically justified, and increasingly required by funders/journals/regulators. He argues for integration of molecular identity into LIMS/FAIR pipelines and for movement from episodic QC to continuous, automated provenance.

 

The manuscript is well-documented and useful to the community.

I request some minor revisions in order to increase rigor and utility.

1) Please improve the transparency of review methods, add method paragraph. The review does not appear to report a reproducible literature search strategy (databases searched, date ranges, inclusion/exclusion criteria, search strings) or a PRISMA flow for included citations. For readers who want to re-run or validate the evidence base, this limits reproducibility.

 

2) Please provide a stronger sensitivity analysis for economic claims. Temper economic certainty by reporting ranges and noting key assumptions. Add a brief prioritized research agenda with specific study designs and metrics.

Author Response

Dear Reviewer 1,

I sincerely thank you for the careful evaluation of our manuscript and the constructive comments, which have helped me to improve the rigor, clarity, and usefulness of the review. Please find attached a pdf file in which I provide a point-by-point to each comment and indicate the corresponding changes made in the revised manuscript.

Regards

Ralf Weiskirchen

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

This is a review that explains why routine STR testing matters and gives useful practical background (standards, databases, workflows). However, some key numbers and interpretations need to be corrected and clarified to avoid contradictions and overstatement.

Major comments

1. Text says ~300 uses (4500), but Fig. 2A says ~3250 studies (48750). Please fix the inconsistency and show the calculation. Also, “falsified data” is too strong unless you are alleging misconduct. Use “potentially unreliable/invalid results” instead.
2. Authors use ANSI/ATCC (≥80% allele concordance; 60–79% partial) but also “≥90% similarity” (Tanabe) as “same cell line.” Please clearly separate these metricss and justify the different cutoffs and don’t present them as equivalent.
3. Authors claim in the abstract >3000% ROI at 0.07% “contamination risk,” but Table 3 assumes 10% misidentification risk/year, €100k damage, 0% discount, and €0 internal cost to get 3233%. Add a short sensitivity check and explain how 0.07% relates to 10% (or state they are not comparable).
4. Numbers like “40% fewer interventions” and “60% fewer false positives,” plus “real-time” RL PCR optimization and near-term blockchain provenance, should be backed with solid evidence and limits, or clearly labeled as future/perspective.

Minor comments

1. Please tone down wording such as “papers worthless” and “falsified data” because it can read as an allegation. Prefer more precise language (misidentification/irreproducibility).
2. Supplementary Materials were not available to me. Please upload them. For the AI-based cost–benefit model, provide the exact prompt and enough details/calculation steps to reproduce the result, and state what was AI-generated vs author-checked

Author Response

Dear Reviewer 2,

I sincerely thank you for the careful evaluation of our manuscript and the constructive comments, which have helped me to improve the rigor, clarity, and usefulness of the review. Please find attached a pdf file in which I provide a point-by-point to each comment and indicate the corresponding changes made in the revised manuscript.

Regards

Ralf Weiskirchen

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

no further comments