National Toxicology Program

National Toxicology Program
https://ntp.niehs.nih.gov/go/iccvam-milestones

Project Milestones

This page summarizes the status of ongoing and completed ICCVAM alternative test method evaluation projects, as well as projects to which NICEATM, ICCVAM, and ICCVAM agency scientists are contributing.

Alternative test methods:

  • Replace animals with non-animal systems or one animal species with a less highly developed one (for example, replacing a mouse with a fish)
  • Reduce the number of animals used to the minimum number required to obtain scientifically valid data
  • Refine procedures to lessen or eliminate animal pain and distress

Acute Systemic Toxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Up-and-Down Procedure (oral) 2001 2001, revised 2008 (OECD TG 425)
Fixed Dose Procedure (oral) (Not applicable - adopted via OECD Test Guideline) 2002 2002 (OECD TG 420)
Acute Toxic Class Method (oral) (Not applicable - adopted via OECD Test Guideline) 2002 2002 (OECD TG 423)
In Vitro Cytotoxicity Test Methods: 3T3 Cells 2008 2010 (OECD GD 129)
In Vitro Cytotoxicity Test Methods: NHK Cells 2008 2010 (OECD GD 129)
Acute Inhalation Toxicity (update to reduce animal use) (Not applicable - adopted via OECD Test Guideline)

2009

2009 (OECD TG 403)
Acute Toxic Class Method (inhalation) (Not applicable - adopted via OECD Test Guideline) 2009 2009 (OECD TG 436)
Fixed Concentration Procedure (inhalation) In progress (OECD) (Not applicable - to be adopted via OECD Test Guideline)   Anticipated 2017 (OECD)

Biologics and Vaccines

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
USDA Policy Concerning the Use of Humane Endpoints in Biologics Testing 2004 2002 (European Pharmacopoeia)
In Vitro Alternative for Pertussis Vaccine Safety Testing In progress      

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Dermal Absorption

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
In Vitro Dermal Absorption (Not applicable - adopted via OECD Test Guideline) 2004 2004 (OECD TG 428)

Dermal Corrosivity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Corrositex® Assay

2000

2006 (OECD TG 435)
EpiDermTM Assay

2004

2004 (OECD TG 431)
EPISKINTM Assay

2004

2004 (OECD TG 431)
Rat Transcutaneal Electrical Resistance Assay

2004

2004 (OECD TG 430)
SkinEthic RHE Assay (Not applicable - adopted via OECD Test Guideline)

2004

2004 (OECD TG 431)
Acute Dermal Corrosion (reduction and refinement of animal use) (Not applicable - adopted via OECD Test Guideline)

2015

2015 (OECD TG 404)
LabCyte EPI-MODEL24 SIT assay (Not applicable - adopted via OECD Test Guideline)

2015

2015 (OECD TG 439)

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Dermal Irritation

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
EpiDermTM Assay (Not applicable - adopted via OECD Test Guideline) 2010 2010 (OECD TG 439)
EPISKINTM Assay (Not applicable - adopted via OECD Test Guideline) 2010 2010 (OECD TG 439)
SkinEthic RHE Assay (Not applicable - adopted via OECD Test Guideline) 2010 2010 (OECD TG 439)
Acute Dermal Corrosion (reduction and refinement of animal use) (Not applicable - adopted via OECD Test Guideline)

2015

2015 (OECD TG 404)
LabCyte EPI-MODEL24 SIT (Not applicable - adopted via OECD Test Guideline)

2015

2015 (OECD TG 439)

Dermal Phototoxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
3T3 NRU Phototoxicity Test (Not applicable - adopted via OECD Test Guideline) 2004 2004 (OECD TG 432)
3T3 NRU Phototoxicity Test: Application to UV Filter Chemicals (Not applicable - adopted via OECD Test Guideline) 2004 2004 (OECD TG 432)

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Developmental Toxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Frog Embryo Teratogenesis Assay: Xenopus (FETAX) ICCVAM evaluation in 2000 found that FETAX was not sufficiently reliable for regulatory use and provided recommendations for improving accuracy and reliability
Combined repeated dose toxicity study with the reproduction/development toxicity screening test (potential for reduced use of animals) (Not applicable - adopted via OECD Test Guideline)

2015

2015 (OECD TG 422)
Hand-1 luciferase embryonic toxicity screen In progress (JaCVAM)        

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Endocrine Disruptor

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
In vitro Androgen Receptor (AR) Binding ICCVAM review in 2004 found validation studies conducted to date to be inadequate - ICCVAM published recommendations on future validation studies
In vitro AR Transcriptional Activation (TA) ICCVAM review in 2004 found validation studies conducted to date to be inadequate - ICCVAM published recommendations on future validation studies
In vitro Estrogen Receptor (ER) Binding ICCVAM review in 2004 found validation studies conducted to date to be inadequate - ICCVAM published recommendations on future validation studies
In vitro ER TA ICCVAM review in 2004 found validation studies conducted to date to be inadequate - ICCVAM published recommendations on future validation studies
H295R steroidogenesis assay (Not applicable - adopted via OECD Test Guideline)

2011

2011 (OECD TG 456)
BG1Luc ER TA Assay Evaluation 2012 (OECD TG 457)
Performance-based test guideline for stably transfected TA in vitro assays to detect estrogen receptor agonists and antagonists (Not applicable - adopted via OECD Test Guideline)

2012

2012 (OECD TG 455)
MCF-7 Cell Proliferation Assay Evaluation Results of NICEATM validation study indicated that the MCF-7 cell proliferation assay protocols require additional development to improve interlaboratory reproducibility
Performance-based test guideline for human recombinant estrogen receptor in vitro assays to detect chemicals with ER binding affinity (Not applicable - adopted via OECD Test Guideline)

2015

2015 (OECD TG 493)
AR-Ecoscreen In progress (JaCVAM)        
Performance-based test guideline for stably transfected TA in vitro assays to detect androgen receptor agonists and antagonists (Not applicable - adopted via OECD Test Guideline)

2016

2015 (OECD TG 458)

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Genetic Toxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
In Vitro Micronucleus Assay (Not applicable - adopted via OECD Test Guideline) 2010 2010 (OECD TG 487)
In Vivo Comet Assay (Not applicable - adopted via OECD Test Guideline) 2014 2014 (OECD TG 489)
In Vitro Comet Assay In progress (JaCVAM)        
In Vitro Syrian Hamster Embryo Cell Transformation Assays (Not applicable - to be adopted via OECD Guidance Document)   Anticipated 2017 (OECD)
BHAS Cell Transformation Assay (Not applicable - to be adopted via OECD Test Guideline)   Anticipated 2017 (OECD)
In vitro mammalian cell gene mutation tests using the thymidine kinase assay (Not applicable - adopted via OECD Test Guideline) 2015 2015 (OECD TG 490)

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Immunotoxicity: Allergic Contact Dermatitis

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Murine Local Lymph Node Assay (LLNA) 1999 2002 (OECD TG 429)
Updated LLNA Protocol (20% animal reduction) 2010 2010 (Updated OECD TG 429)
Reduced LLNA Test Method 2010 2010 (Updated OECD TG 429)
Development of LLNA Performance Standards 2010 2010 (Updated OECD TG 429)
Nonradioactive LLNA Method: LLNA:DA 2011 2010 (OECD TG 442A)
Nonradioactive LLNA Method: LLNA:BrdU-ELISA 2011 2010 (OECD TG 442B)
Nonradioactive LLNA Method: LLNA:BrdU-Flow Cytometry Interlaboratory validation study required        
Use of the LLNA for Testing Pesticide Formulations, Metals, Substances in Aqueous Solutions, and Other Products 2011 2010 (Updated OECD TG 429)
Use of the LLNA for Skin Sensitization Potency Categorization 2009 (UN GHS)
KeratinoSens Assay (Not applicable - adopted via OECD Test Guideline) 2015 2015 (OECD TG 442D)
Direct Peptide Reactivity Assay (Not applicable - adopted via OECD Test Guideline) 2015 2015 (OECD TG 442C)
Human Cell Line Activation Test (Not applicable - adopted via OECD Test Guideline) 2016 2016 (OECD TG 442E)
Myeloid U937 Skin Sensitization Test EURL ECVAM review found that additional development was required to improve reproducibility.
IL-8 Luciferase Assay In progress (JaCVAM)      
Vitrigel-SST Assay In progress (JaCVAM)      
Electrophilic Allergen Screening Assay In progress        
Amino Acid Derivative Reactivity Assay In progress (JaCVAM)        
Nonradioactive LLNA BrdU-Flow Cytometry Method In progress (OECD evaluation of a KoCVAM-sponsored study)      

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Metabolism and Toxicokinetics

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Provision of a Standard for Human Hepatic Metabolism and Toxicity by Assessing as an Indicator Biotransformation Enzyme Induction Using HepaRG® Cells and Cryopreserved Human Hepatocytes In progress (OECD) (Not applicable - to be adopted via OECD Test Guideline)    

Ocular Toxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Bovine Corneal Opacity and Permeability (BCOP) - Ocular Corrosivity/Severe Irritation 2008 2009 (OECD TG 437)
Isolated Chicken Eye (ICE) - Ocular Corrosivity/Severe Irritation 2008 2009 (OECD TG 438)
Hen's Egg Test/Choriallantoic Membrane (HET-CAM) - Ocular Corrosivity/Severe Irritation ICCVAM review in 2007 found that this test method was not sufficiently accurate for regulatory use and recommended additional studies
Isolated Rabbit Eye (IRE) - Ocular Corrosivity/Severe Irritation ICCVAM review in 2007 found that this test method was not sufficiently accurate for regulatory use and recommended additional studies
Integrated Non-animal Testing Strategy for Eye Irritation Potential of Antimicrobial Cleaning Products ICCVAM review in 2009 concluded that data on the testing strategy were not sufficient to demonstrate its utility for the proposed application and recommended additional studies
BCOP - Nonsevere Ocular Irritation ICCVAM review in 2009 found that this test method was not sufficiently accurate for the proposed use and recommended additional studies
HET-CAM - Nonsevere Ocular Irritation ICCVAM review in 2009 found that this test method was not sufficiently accurate for the proposed use and recommended additional studies
ICE - Nonsevere Ocular Irritation ICCVAM review in 2009 found that this test method was not sufficiently accurate for the proposed use and recommended additional studies
IRE - Nonsevere Ocular Irritation ICCVAM review in 2009 found that this test method was not sufficiently accurate for the proposed use and recommended additional studies
Cytosensor Microphysiometer Test Method 2011 In progress (OECD)
Routine use of topical anesthestics, systemic analgesics, and humane endpoints in in vivo testing 2011 2012 (Updated OECD TG 405)
In vivo low volume eye test ICCVAM reviewed the LVET in 2009 and does not recommend its use for prospective ocular safety testing
Fluorescein leakage test method - ocular corrosivity/severe irritation (Not applicable - adopted via OECD Test Guideline) 2012 2012 (OECD TG 460)
Neutral red release test method ICCVAM review in 2009 found that this test method was not adequately validated for regulatory use
Red blood cell haemolysis test method ICCVAM review in 2009 found that this test method was not adequately validated for regulatory use
Short time exposure test (Not applicable - adopted via OECD Test Guideline) 2015 2015 (OECD TG 491)
OptiSafe test method In progress        
Reconstructed human cornea-like epithelium test (EpiOcular) (Not applicable - adopted via OECD Test Guideline) 2015 2015 (OECD TG 492)
SIRC-CVS test method In progress (JaCVAM)      
Vitrigel-EIT In progress (JaCVAM)        

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Pyrogenicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Human Whole Blood/IL-1 In Vitro Pyrogen Test 2009 2010 (European Pharmacopoeia)
Human Whole Blood/ IL-1 In Vitro Pyrogen Test Using Cryopreserved Human Whole Blood 2009 2010 (European Pharmacopoeia)
Human Whole Blood/IL-6 In Vitro Pyrogen Test (WB/IL-6) 2009 2010 (European Pharmacopoeia)
In Vitro Pyrogen Test Using Human Peripheral Blood Mononuclear Cells (PBMC/IL-6) 2009 2010 (European Pharmacopoeia)
Alternative In Vitro Pyrogen Test Using the Human Monocytoid Cell Line MONO MAC 6 (MM6/IL-6) 2009 2010 (European Pharmacopoeia)

Reproductive and Developmental Toxicity

Test Method R&D Validation Studies Evaluation Recommendations
to U.S. Agencies
U.S.
Acceptance
International
Acceptance
Hand-1 Luciferase Assay In progress (JaCVAM)        

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