Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8551680 | Regulatory Toxicology and Pharmacology | 2018 | 36 Pages |
Abstract
With the increasing need to develop in vitro assays to replace animal use, human stem cell-derived methods are emerging and showing outstanding contributions to the toxicological screening of substances. Adult human stem cells such as adipose-derived stem cells (ADSC) and periodontal ligament stem cells (PDLSC) were used as cell substrates for a cytotoxicity assay and toxicity prediction using the neutral red uptake (NRU) assay. First, primary cell cultures from three independent donors, from each tissue source, were characterized as mesenchymal stem cells (MSC) by plastic adherence and appropriate immunophenotype for MSC markers (positive for CD90, CD73, and CD105 and negative for CD11b, CD34, CD45, HLADR, and CD19). Furthermore, ADSC and PDLSC were able to differentiate into adipocytes and osteoblasts when maintained under the same culture conditions previously established for the NRU assay. NRU assays for three reference test substances were performed. R2 was higher than 0.85 for all conditions, showing the feasibility to calculate IC50 values. The IC50 values were then used to predict the LD50 of the test substances, which were comparable to previous results and the ICCVAM standard test report. Primary ADSC and PDLSC showed the potential to be considered as additional models for use in cytotoxicity assays.
Keywords
GHSEURL-ECVAMIMDMD-PBSBMSCOECDMSCICCVAMToxCastFBSDMEMPBSNHKEPAADSCDulbecco's modified Eagle's mediumIscove's modified Dulbecco's mediumNRUEmbryonic stem cell testEuropean Union Reference Laboratory for Alternatives to Animal TestingUnited States Environmental Protection AgencyESTneutral red uptakeOrganization for Economic Co-operation and Developmentfetal bovine serumBone marrow-derived stem cellsAdipose-derived stem cellsPeriodontal ligament stem cellsMesenchymal stem cellsCytotoxicityglobally harmonized system of classification and labeling of chemicalsphosphate-buffered saline solutionInteragency Coordinating Committee on the Validation of Alternative Methods
Related Topics
Life Sciences
Environmental Science
Health, Toxicology and Mutagenesis
Authors
Natássia Caroline Resende Corrêa, Crisciele Kuligovski, Ariane Caroline Campos Paschoal, Ana Paula Ressetti Abud, Carmen Lucia Kuniyoshi Rebelatto, Lidiane Maria Boldrini Leite, Alexandra Cristina Senegaglia, Bruno Dallagiovanna,