ISO INTERNATIONAL STANDARD 19007 First edition 2018-04 Nanotechnologies In vitro MTs assay for measuring the cytotoxic effect of nanoparticles Nanotechnologies - Analyse du MTS in vitro pour la mesure de I'effet cytotoxique des nanoparticules Reference number IS0 19007:2018(E) so Copyright International Organization for Standardization @IS02018 IS0 19007:2018(E) COPYRIGHTPROTECTEDDOCUMENT @ IS0:2018 All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior written permission. Permission can be requested from either IsO at the address below or Iso's member body in the country of the requester. ISO copyright office CP401·Ch.deBlandonnet8 CH-1214 Vernier, Geneva Phone:+4122 749 0111 Fax: +41 22 749 09 47 Email: copyright@iso.org Website: www.iso.org Published in Switzerland Copyightilerational Organizationfor Standardization @ IS0 2018 - All rights reserved IS0 19007:2018(E) Contents Page Foreword ..iv Introduction. ..V 1 Scope. .1 2 Normative references ..1 3 Terms and definitions ..1 4 Symbols and abbreviated terms ..2 5 Materials .3 5.1 Cell line .3 5.2 Assay. .3 5.3 Controls .3 6 Apparatus .4 7 Nanoparticle test sample preparation .4 8 Preparations .5 8.1 General .5 8.2 Culture medium .5 8.3 Preparation of cell stock culture .5 8.4 Verifyviablecell growth .6 8.5 Verification of plate reader uniformity .6 8.6 Control preparation 6 8.6.1 Control description.. .6 8.6.2 CdsO4 stock solution preparation (10mM) 7 8.6.3 Nanoparticle control suspension preparation .7 8.7 Precision pipetting. 9 Characterization of nanoparticle impact on cell viability .7 9.1 General .7 9.2 Preparation of the cell plate 9.3 Prepare the nanoparticle dosing plate .9 9.4 Expose cells to nanoparticles in culture medium 11 9.5 Expose cells to MTS Assay .11 9.6 Measurement of formazan absorbance .12 10 Cell viability analysis. ..12 11 Interpertation of Assay Results ..12 Annex A (informative) Potential cell lines and assays .13 Annex B (informative) Example: the MTS assay using the A549 cell line (EMPA-NIST protocol) ....14. Annex C (informative) Example: MTS assay using the RAW 264.7 cell line (IANH protocol) .23 Bibliography .31 CopyightIntemational OrgazArights reserved iii