Service interruption on Monday 11 July from 12:30 to 13:00: all the sites of the CCSD (HAL, EpiSciences, SciencesConf, AureHAL) will be inaccessible (network hardware connection).
Skip to Main content Skip to Navigation
Journal articles

Key parameter optimization using multivariable linear model for the evaluation of the in vitro estrogenic activity assay in T47D cell lines (CXCL‐test)

Abstract : In comparison to analytical tools, bioassays provide higher sensitivity and more complex evaluation of environmental samples and are indispensable tools for monitoring increasing in anthropogenic pollution. Nevertheless, the disadvantage in cellular assays stems from the material variability used within the assays, and an interlaboratory adaptation does not usually lead to satisfactory test sensitivities. The aim of this study was to evaluate the influence of material variability on CXCL12 secretion by T47D cells, the outcome of an estrogenic activity assay, the CXCL-test. For this purpose, the cell line sources, sera suppliers, experimental and seeding media, and the amount of cell/well were tested. The multivariable linear model (MLM), employed as an innovative approach in this field for parameter evaluation, identified that all the tested parameters had significant effects. Knowledge of the contributions of each parameter has permitted step-by-step optimization. The most beneficial approach was seeding 20,000 cells/well directly in treatment medium and using DMEM for the treatment. Great differences in both basal and maximal cytokine secretions among the three tested cell lines and different impacts of each serum were also observed. Altogether, both these biologically based and highly variable inputs were additionally assessed by MLM and a subsequent two-step evaluation, which revealed a lower variability and satisfactory reproducibility of the test. This analysis showed that not only parameter and procedure optimization but also the evaluation methodology must be considered from the perspective of interlaboratory method adaptation. This overall methodology could be applied to all bioanalytical methods for fast multiparameter and accurate analysis.
Document type :
Journal articles
Complete list of metadata

https://hal-anses.archives-ouvertes.fr/anses-03512680
Contributor : Emmanuelle Chiffoleau Connect in order to contact the contributor
Submitted on : Wednesday, January 26, 2022 - 11:31:21 AM
Last modification on : Thursday, June 23, 2022 - 2:27:50 PM
Long-term archiving on: : Wednesday, April 27, 2022 - 6:38:29 PM

File

Linhartova et al-2021-Key para...
Files produced by the author(s)

Licence


Distributed under a Creative Commons Attribution - NonCommercial 4.0 International License

Identifiers

Citation

Lucie Linhartova, Nathalie Costet, Farzad Pakdel, Tomas Cajthaml, Denis Habauzit. Key parameter optimization using multivariable linear model for the evaluation of the in vitro estrogenic activity assay in T47D cell lines (CXCL‐test). Journal of Applied Toxicology, Wiley, 2022, 42 (7), pp.1121-1136. ⟨10.1002/jat.4280⟩. ⟨anses-03512680⟩

Share

Metrics

Record views

33

Files downloads

14