Name of the deliverable

Lead Beneficiary

SOP - Strategy - Guidance document


Report on identification and setting of categorization, read-across, and extra/interpolation criteria




Report on the development of a solubility testing procedure




In vitro screening methodology for absorption or crossing of other barriers


SOPs available in deliverable:

  • SOP 01 Protocol for organotypic cell culture modified for exposure to nanomaterials
  • SOP 02 Caco-2 cell culture and differentiation
  • SOP 03 Evaluation of NMs impact on Caco-2 cell barrier model


In vitro screening methodology to evaluate toxicity by inhalation




Report on cell type and in vitro-in vivo correlation studies for inhalation toxicity




Identification and optimization of the most suitable in vitro methodology


SOPs available in deliverable:

  • SOP 01 LAL Assay for Nanoparticles
  • SOP 02 LDH assay
  • SOP 03 Reaction Oxygen Species Detection
  • SOP 04 Human Lung Cell transformation assay. Long term chronic experiment
  • SOP 05 Apoptosis/Necrosis Analysis
  • SOP 06 Production of pro-inflammatory cytokines


Develop a rapid high throughput screening methodology to evaluate NM toxicity



      SOP 01 SOP for Dispersion
      SOP 02 Real-time label-free impedance-based nanotoxicity assessment
      SOP 03 SOP for High Throughput Cell Impedance Measurement
      SOP 04 Label-free nanotoxicity assessment by impedance-based flow cytometry
      SOP 05 Viability protocol by using a Cell Counter
      SOP 06 HEL16T008 AlamarBlue Assay
      SOP 07 HEL17T010 Colony forming efficiency assay
      SOP 08 Cytotoxicity by MTS - reference to NanoValid protocol
      SOP 09 TaqMan real-time Reverse Transcription PCR
      SOP 10 HEL11T005 HTS Comet Assay with and without FPG - 12 wells
      SOP 11 HTS Comet Assay with and without FPG - 20 wells
      SOP 12 HEL11T007 Mammalian in vitro HPRT Mutation test
      SOP 13 HEL16T009 Mouse Lymphoma Assay in vitro
      SOP 14 Micronucleus Assay
      SOP 15 Micronucleus assay using Flow Cytometry
      SOP 16 High Content Analysis-based nanotoxicity assessment
      SOP 17 Ion Beam Microscopy
      SOP 18 Confocal Raman Mcrospectroscopy
      SOP 19 Whole blood cell assay for cytokine production


Decision tree for risk assessment of MNMs


RIVM FlowChart NANoREG nanospecific risk assessment strategy

D5.09 Summary report WP5    

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Information on “commercial use” and attribution can be found here.