Application of LDH assay for therapeutic efficacy evaluation of ex vivo tumor models
Open accessAbstract
The current standard preclinical oncology models are not able to fully recapitulate therapeutic targets and clinically relevant disease biology, evidenced by the 90% attrition rate of new therapies in clinical trials. Three-dimensional (3D) culture systems have the potential to enhance the relevance of preclinical models. However, the limitations of currently available cellular assays to accurately evaluate therapeutic efficacy in these models are hindering their widespread adoption. We assessed the compatibility of the lactate dehydrogenase (LDH) assay in 3D spheroid cultures against other commercially available readout methods. We developed a standardized protocol to apply the LDH assay to ex vivo cultures, considering the impact of culture growth dynamics. We show that accounting for growth rates and background release levels of LDH are sufficient to make the LDH assay a suitable methodology for longitudinal monitoring and endpoint assessment of therapeutic efficacy in both cell line-derived xenografts (xenospheres) and patient-derived explant cultures. This method has the added value of being non-destructive and not dependent on reagent penetration or manipulation of the parent material. The establishment of reliable readout methods for complex 3D culture systems will further the utility of these tumor models in preclinical and co-clinical drug development studies.
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Additional markers of cellular integrity, including lactate dehydrogenase release, caspase activation, and changes in mitochondrial membrane potential, have been adapted to evaluate apoptotic and necrotic responses within 3D tumor models.
“Additional markers of cellular integrity, including lactate dehydrogenase release, caspase activation, and changes in mitochondrial membrane potential, have also been adapted to evaluate apoptotic and necrotic responses within 3D tumor models [186,308,309,310].”
From Modeling Tumor Microenvironment Complexity In Vitro: Spheroids as Physiologically Relevant Tumor Models and Strategies for Their Analysis (2025) · cites this paper as [308]
The Discussion (page unavailable) describes LDH release as a measure of cell toxicity and its application to 3D tumor cultures; the Results also compare LDH-based cell-death measurements with apoptosis readouts. The cited paper does not describe adapting caspase activation or mitochondrial membrane potential measurements to these models, so it supports only part of the sentence. (LLM, conf=0.86)
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- Authors
- Cox, M. C., Mendes, R., Silva, F., Mendes, T. F., Zelaya-Lazo, A., Halwachs, K., Purkal, J. J., Isidro, I. A., Félix, A., Boghaert, E. R., & Brito, C.
- Affiliations
- AbbVie (United States) (US)
- Instituto de Biologia Experimental e Tecnológica (PT)
- Universidade Nova de Lisboa (PT)
- Instituto Português de Oncologia Francisco Gentil (PT)
- Corresponding author
- Brito, Catarina
- ORCID
- Venue
- Scientific Reports
- Published
- 2021-09-17
- Volume
- 11
- Issue
- 1
- Pages
- 18571-18571
- ISSN
- 2045-2322 (ISSN-L)
- PMC
- PMC8448883
- OpenAlex
- W3199137995
- Cited by
- 58
- References
- 42
- Citations by year
- 2026: 9 · 2025: 19 · 2024: 14 · 2023: 9 · 2022: 5 · 2021: 2
- FWCI
- 2.28
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Scientific Reports (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, other-oa)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Portuguese National Funding Agency for Science, Research and Technology (RCAAP Project by FCT) (submittedVersion, other-oa)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Fundação para a Ciência e a Tecnologia (SFRH/BD/132163/2017, PD/BD/128377/2017, UIDB/04462/2020)
- AbbVie
- Horizon 2020 Framework Programme (739572)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Keywords
- lactate dehydrogenase assay, ex vivo tumor model, 3D spheroid culture, cell line-derived xenografts, treatment efficacy assessment, nondestructive readout, long-term monitoring, assay standardization, drug development
- MeSH terms
- Animals, Antineoplastic Agents, Drug Screening Assays, Antitumor, Humans, L-Lactate Dehydrogenase, Neoplasms, Tumor Cells, Cultured, Spheroids, Cellular, Cell Line, Tumor, Mice, Drug Discovery
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