Update on JaCVAM | Japanese Center for the Validation of Alternative Methods, National Institute of Health Science

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Classification	Test method	Validation study	Peer reviews by experts	Protocol	Test guidelines	Recommendation to government
01 Corrosivity Test	01 Transcutaneous Electrical Resistance Test Method (TER)	EURL ECVAM The ECVAM International Validation Study on In Vitro Tests for Skin Corrosivity. 2. Results and Evaluation by the Management Team A Prevalidation Study on In Vitro Skin Corrosivity Testing	EURL ECVAM & ICCVAM EPISKIN, EpiDerm, and Rat Skin Transcutaneous Electrical Resistance Assay STATEMENT ON THE SCIENTIFIC VALIDITY OF THE RAT SKIN TRANSCUTANEOUS ELECTRICAL RESISTANCE (TER) TEST		OECD TG No. 430	Oct.2017 JaCVAM statement Evaluation report on an Alternative Method for Assessing Skin Corrosion Potential Using Transcutaneous Electrical Resistance(TER)
	02 Reconstructed Human Epidermis (RHE) Test Method, EpiSkin™, EpiDerm™ SCT, SkinEthics™ RHE, epiCS®	EURL ECVAM The ECVAM International Validation Study on In Vitro Tests for Skin Corrosivity. 2. Results and Evaluation by the Management Team The ECVAM Prevalidation Study on the Use of EpiDerm for Skin Corrosivity Testing. Adaptation of the validated SkinEthic™ Reconstructed Human Epidermis (RHE) skin corrosion test method to 0.5 cm2 tissue sample Epidermal-skin-test 1000 (EST-1000)—A new reconstructed epidermis for in vitro skin corrosivity testing	EURL ECVAM & ICCVAM EPISKIN, EpiDerm, and Rat Skin Transcutaneous Electrical Resistance Assay STATEMENT ON THE SCIENTIFIC VALIDITY OF THE EPISKIN^TM TEST (AN IN VITRO TEST FOR SKIN CORROSIVITY) ESAC STATEMENT ON THE SCIENTIFIC VALIDITY OF AN IN-VITRO TEST METHOD FOR SKIN CORROSIVITY TESTING ESAC Opinion on the SkinEthic™ Human Corneal Epithelium (HCE) Eye Irritation Test (EIT)	RAW MATERIALS / VALIDATED TEST METHOD (under OECD Test Guideline 431) DB-ALM Protocol n° 119 : EpiDerm^TM Skin Corrosivity Test DB-ALM Protocol n° 118 : EpiSkin^TM Skin Corrosivity Test epiCS®	OECD TG No. 431	Jun.2017 JaCVAM statement Evaluation report on an Alternative Method for Assessing Skin Corrosion Potential Using Reconstructed Human Epidermis(RHE)
	03 Reconstructed Human Epidermis (RHE) Test Method, LabCyte EPI- MODEL24 SCT	JaCVAM	JaCVAM		OECD TG No. 431	Dec.2021 JaCVAM statement
	04 In vitro Membrane Barrier Test Method	EURL ECVAM	EURL ECVAM & ICCVAM STATEMENT ON THE APPLICATION OF THE CORROSITEX® ASSAY FOR SKIN CORROSIVITY TESTING Corrositex®:An In Vitro Test Method for Assessing Dermal Corrosivity Potential of Chemicals		OECD TG No. 435	Oct.2017 JaCVAM statement Evaluation report on an Alternative Method for Assessing Skin Corrosion Potential Using In Vitro Membrane Barrier
	05 Reconstructed Human Epidermis (RHE) Test Method, Vitrolife-skin^TM	JSAAE	JaCVAM			Aug.2008 JaCVAM statement Evaluation report on the Vitrolife-Skin^TM, a 3-dimensional cultured skin model for skin corrosivity testing

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Classification	Test method	Validation study	Peer reviews by experts	Protocol	Test guidelines	Recommendation to government
02 Skin Irritation Test	01 Reconstructed Human Epidermis (RHE) Test Method, EpiSkin^TM, EpiDerm^TM SIT, SkinEthics^TM RHE	EURL ECVAM	EURL ECVAM STATEMENT ON THE SCIENTIFIC VALIDITY OF THE EPISKIN^TM TEST (AN IN VITRO TEST FOR SKIN CORROSIVITY) ESAC Peer review of submission of the follow-up validation study of the Modified EpiDerm^TM Skin Irritation Test (SIT) for hazard identification and labelling of chemicals according to EU classification system. ESAC Peer review of submission of external catch up validation study of In vitro Reconstructed Human Epidermis (RHE) assay for Skin Irritation Testing REPORTS OF THE PEER REVIEWS OF THE epiCS AND SKIN+ TEST METHODS IN VIEW OF THEIR INCLUSION IN TEST GUIDELINE 439 ON IN VITRO SKIN IRRITATION	EpiDerm EpiSkin SkiEthics EpiCS	OECD TG No. 439	Jan.2013 JaCVAM statement (EPISKIN) Evaluation report on the in-vitro tests for skin irritation JaCVAM statement (EpiDerm and SkinEthics) Evaluation report on the two in-vitro assays for skin irritation testing
	02 Reconstructed Human Epidermis (RHE) Test Method, LabCyte EPI-MODEL24 SIT	JaCVAM	OECD		OECD TG No. 439	Nov.2013 JaCVAM statement Evaluation report on in vitro assay using the reconstructed human epidermis models, LabCyte EPI-MODEL24 for skin irritation testing
	03 Reconstructed Human Skin Test, LbL-3D Skin SIT	JaCVAM
	04 Reconstructed Human Epidermis (RHE) Test Method, LabCyte EPI-MODEL24 SIT for Medical Devices	NIHS Div.of Medical Divices/JaCVAM LabCyte_EPI-MODEL24_RRS_Report Appendixes		SOP of LabCyte EPI-MODEL24 Skin Irritation test method (LabCyte EPI-MODEL24 SIT) for medical device

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Classification	Test method	Validation study	Peer reviews by experts	Protocol	Test guidelines	Recommendation to government
03 Phototoxicity Test	01 In vitro 3T3 NRU Phototoxicity Test	Cosmetic Europe/ EURL ECVAM			OECD TG No. 432
	02 ROS (Reactive Oxygen Species) Assay for Photoreactivity	JaCVAM Validation report for the international validation study on ROS (Reactive Oxygen Species) assay as a test evaluating phototoxic potential of chemicals (Atlas Suntest version) Validation report for the international validation study on ROS (Reactive Oxygen Species) assay as a test evaluating phototoxic potential of chemicals (Seric version) Validation report for the international validation study on ROS (Reactive Oxygen Species) assay as a test evaluating phototoxic potential of chemicals (Atlas Suntest version)	JaCVAM	最新版 03_02_E3.pdf 旧版 03_02_E1.pdf 03_02_E2.pdf	ICH S10 OECD TG No. 495 GD	Jan.2016 (in revision) JaCVAM statement Evaluation Report on an Alternative Test Method for Photosafety: Reactive Oxygen Species Assay
	03 Yeast-RBC (Red Blood Cell) Assay	JSAAE	JaCVAM			Evaluation Report on battery system with yeast growth inhibition assay and red blood cells hemolysis assay to predict phototoxicity of chemicals
	04 Reconstructed Human Epidermis (RHE) Test Method	BfR	EURL ECVAM		OECD TG No. 498

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Classification	Test method	Validation study	Peer reviews by experts	Protocol	Test guidelines	Recommendation to government
04 Eye Irritation Test	01 OECD TG No.405 (2012)		ICCVAM		OECD TG No. 405	Jan.2014 JaCVAM statement Evaluation Report on the Revised OECD Test Guideline for Eye irritation/Corrosion(TG405)
	02 Bovine Corneal Opacity and Permeability Test Method (BCOP)	EURL ECVAM & ICCVAM	EURL ECVAM & ICCVAM ESAC STATEMENT ON THE CONCLUSIONS OF THE ICCVAM RETROSPECTIVE STUDY ON ORGANOTYPIC IN VITRO ASSAYS AS SCREENING TESTS TO IDENTIFY POTENTIAL OCULAR CORROSIVES AND SEVERE IRRITANTS AS DETERMINED BY US EPA, EU(R41) AND UN GHS CLASSIFICATIONS IN A TIERED TESTING STRATEGY, AS PART OF A WEIGHT OF EVIDENCE APPROACH Expert Panel Evaluation of the Current Validation Status of In Vitro Test Methods for Identifying Ocular Corrosives and Severe Irritants Expert Panel Report: Evaluation of the Current Validation Status of In Vitro Test Methods for Identifying Ocular Corrosives and Severe Irritants – Addendum STREAMLINED SUMMARY DOCUMENT SUPPORTING OECD GUIDELINE 437 ON THE BOVINE CORNEAL OPACITY AND PERMEABILITY FOR EYE IRRITATION/CORROSION	ICCVAM-Recommended Protocol for Using the Bovine Corneal Opacity and Permeability (BCOP) Test Method DB-ALM Protocol n° 124 : Bovine Corneal Opacity and Permeability (BCOP) Assay - SOP of Microbiological Associates LTD., UK	OECD TG No. 437	Jan.2014 JaCVAM statement Evaluation report on the in vitroocular toxicity test methods for identifying ocular corrosive and severe irritants: Bovine Corneal Opacity and Permeability Test Method JaCVAM statement (OECD TG revised NO.437) Evaluation report on the Bovine Corneal Opacity and Permeability Test (BCOP) for alternative to eye irritation test
	03 Isolated Chicken Eye Test Method	EURL ECVAM & ICCVAM	EURL ECVAM & ICCVAM ESAC STATEMENT ON THE CONCLUSIONS OF THE ICCVAM RETROSPECTIVE STUDY ON ORGANOTYPIC IN VITRO ASSAYS AS SCREENING TESTS TO IDENTIFY POTENTIAL OCULAR CORROSIVES AND SEVERE IRRITANTS AS DETERMINED BY US EPA, EU(R41) AND UN GHS CLASSIFICATIONS IN A TIERED TESTING STRATEGY, AS PART OF A WEIGHT OF EVIDENCE APPROACH Expert Panel Evaluation of the Current Validation Status of In Vitro Test Methods for Identifying Ocular Corrosives and Severe Irritants Expert Panel Report: Evaluation of the Current Validation Status of In Vitro Test Methods for Identifying Ocular Corrosives and Severe Irritants – Addendum Streamlined Summary Document Supporting OECD Test Guideline 438 on the Isolated Chicken Eye for Eye Irritation/Corrosion	ICCVAM-Recommended Protocol for Using the Isolated Chicken Eye (ICE) Test Method DB-ALM Protocol n° 80 : Chicken Enucleated Eye Test (CEET) / Isolated Chicken Eye (ICE) test	OECD TG No. 438	Nov.2019 JaCVAM statement Evaluation report on the in vitroocular toxicity test methods for identifying ocular corrosive and severe irritants: Isolated Chicken Eye Test Method Evaluation report on the Isolated Chicken Eye Test (ICE) for alternative to eye irritation test
	04 Fluorescein Leakage (FL)Test Method	EURL ECVAM & ICCVAM	EURL ECVAM PEER REVIEW PANEL REPORT ON THE RETROSPECTIVE VALIDATION OF CYTOTOXICITY/CELL-FUNCTION BASED IN VITRO ASSAYS (EYE IRRITATION) STATEMENT ON THE SCIENTIFIC VALIDITY OF CYTOTOXICITY/CELL-FUNCTION BASED IN VITRO ASSAYS FOR EYE IRRITATION TESTING STREAMLINED SUMMARY DOCUMENT RELATED TO THE FLUORESCEIN LEAKAGE (FL) TEST METHOD FOR IDENTIFICATION OF OCULAR CORROSIVES AND SEVERE IRRITANTS (TG 460)		OECD TG No. 460	Jan.2013 JaCVAM statement Evaluation Report on the Fluorescein Leakage(FL) test for eye irritation testing
	05 Short Time Exposure in vitro Test Method	JaCVAM	ICCVAM		OECD TG No. 491	Mar.2016 JaCVAM statement Evaluation Report on the Alternative Test Method for Ocular Irritation: Short Time Exposure in vitro Test Method
	06 Reconstructed Human Cornea-like Epithelium(RhCE) Test Method, EpiOcular^TM EIT、SkinEthics^TM HCE EIT	Cosmetic Europe/ EURL ECVAM Cosmetics Europe multi-laboratory pre-validation of the EpiOcular™ reconstituted human tissue test method for the prediction of eye irritation Cosmetics Europe Multi-Laboratory Pre-Validation of the SkinEthic™ Reconstituted Human Corneal Epithelium Test Method for the Prediction of Eye Irritation.	EURL ECVAM ESAC Opinion on the EURL ECVAM Eye Irritation Validation Study (EIVS) on EpiOcular™ EIT and SkinEthic™ HCE and a related Cosmetics Europe study on HPLC/UPLC-spectrophotometry as an alternative endpoint detection system for MTT-formazan ESAC OPINION based on the ESAC Scientific Peer Review of the ECVAM Eye Irritation Validation Study (EIVS) and a related Cosmetics Europe study on HPLC/UPLC-photometry as an alternative endpoint detection system for formazan reaction product	DB-ALM Protocol n° 191: SkinEthic™ HCE Eye Irritation Test Solid (EITS) DB-ALM Protocol n° 190 : SkinEthic™ HCE Eye Irritation Test Liquid (EITL)	OECD TG No. 492	Mar.2018 JaCVAM statement (EpiOcular^TM EIT) Evaluation Report on the Human Cornea-like Epithelium Test Method JaCVAM statement (SkinEthics^TM HCE EIT) Evaluation Report on a Reconstructed Human Cornea-Like Epithelium Test Method: The SkinEthic^TM Human Corneal Epithelium Eye Irritation Test
	07 Reconstructed Human Cornea-like Epithelium(RhCE) Test Method, LabCyte CORNEA-MODEL24 EIT	JaCVAM VALIDATION STUDY FOR LabCyte CORNEA-MODEL24 EYE IRRITATION TEST New Established In Vitro Eye Irritation Test Method using Reconstructed Human Corneal Epithelium, LabCyte CORNEA-MODEL24	JaCVAM			Feb.2019 JaCVAM statement
	08 Reconstructed Human Cornea-like Epithelium(RhCE) Test Method, MCTT	KoCVAM	OECD
	09 Reconstructed Human Cornea-like Epithelium(RhCE) Test Method,SkinEthicsTM HCE TTT	L’Oreal	Independent peer reviewer		OECD TG No.492B	Feb.2024 JaCVAM statement
	10 Vitrigel-EIT (Eye Irritancy Test Method)	JaCVAM Validation Study of the Vitrigel-EIT method as an alternative to in vivo eye irritation testing Study plan for the validation trial on Vitrigel-EIT as alternative to eye irritation testing Appendix 8.13 Predictive capacity of the Vitrigel-EIT for 114 chemicals considering DRD Appendix 8.8 Predictive capacity of the Vitrigel-EIT for 114 chemicals Multi-laboratory Validation Study of the Vitrigel-Eye Irritancy Test Method as an Alternative to In Vivo Eye Irritation Testing.	JaCVAM	【Latest version】 Ver1.82j Standard Protocol for the Vitrigel-EIT method 【Old version】 Standard Protocol for the Vitrigel-EIT method	OECD TG No. 494	Jun.2022 JaCVAM statement
	11 Cytosensor Microphysiometer (CM) Toxicity Test Method	EURL ECVAM Validation of the cytosensor for in vitro cytotoxicity studies Validation of the Cytosensor™ Microphysiometer for In Vitro Cytotoxicity Testing	EURL ECVAM Independent Scientific Peer Review Panel Report: Evaluation of the Validation Status of Alternative Ocular Safety Testing Methods and Approaches STATEMENT ON THE SCIENTIFIC VALIDITY OF CYTOTOXICITY/CELL-FUNCTION BASED IN VITRO ASSAYS FOR EYE IRRITATION TESTING		OECD
	12 Ocular Irritection (OI®)	InVitro International	EURL ECVAM		OECD TG No. 496
	13 OptiSafe Eye Irritation Test™ (OS)	InVitro International	EURL ECVAM		OECD TG No. 496
	14 SIRC-CVS (Crystal Violete Staining):TEA Assay	JaCVAM Validation Study for the Statens Seruminstitut Rabbit Cornea–Crystal Violet Staining Cytotoxicity Test Method with Triethanolamine (SIRC-CVS:TEA Test Method) Study Report Version 9.0 Appendices	JaCVAM			Jul.2021 JaCVAM statement
	15 Reconstructed Human Epidermis (RHE) Test Method, MATREX	JaCVAM
	16 Defined Approach for for Serious Eye Damage/Eye Irritation				OECD TG No.467

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Classification	Test method	Validation study	Peer reviews by experts	Protocol	Test guidelines	Recommendation to government
05 Skin Sensitization Test	01 LLNA	EURL ECVAM	ICCVAM ICCVAM Test Method Evaluation Report The Reduced Murine Local Lymph Node Assay: An Alternative Test Method Using Fewer Animals to Assess the Allergic Contact Dermatitis Potential of Chemicals and Products ICCVAM Test Method Evaluation Report: Usefulness and Limitations of the Murine Local Lymph Node Assay for Potency Categorization of Chemicals Causing Allergic Contact Dermatitis in Humans ESAC Statement on the Reduced Local Lymph Node Assay (rLLNA)		OECD TG No. 429
	02 rLLNA	EURL ECVAM & ICCVAM	ICCVAM		OECD TG No. 429	Jan.2013 JaCVAM statement Evaluation report on the reduced Local Lymph Node Assay (rLLNA)
	03 LLNA:DA	JSAAE Validation studies on an alternative endpoint for the local lymph node assay (LLNA-DA): Importance of study management Interlaboratory validation of the modified murine local lymph node assay based on adenosine triphosphate measurement	JaCVAM or ICCVAM The Reduced Murine Local Lymph Node Assay: An Alternative Test Method Using Fewer Animals to Assess the Allergic Contact Dermatitis Potential of Chemicals and Products ESAC Statement on the Reduced Local Lymph Node Assay (rLLNA)		OECD TG No. 442A	Jan.2013 JaCVAM statement (2008) First evaluation report on the LLNA:DA for skin sensitization testing JaCVAM statement (2013) Evaluation report on the Local Lymph Node Assay (LLNA):DA for skin sensitization assay
	04 LLNA:BrdU-ELISA	JSAAE Inter‐laboratory validation of the modified murine local lymph node assay based on 5‐bromo‐2′‐deoxyuridine incorporation	JaCVAM or ICCVAM	ICCVAM-Recommended Test Method Protocol: The Murine Local Lymph Node Assay: 2-Bromodeoxyuridine-ELISA Test Method (LLNA: BrdU-ELISA), a Nonradioactive Alternative Test Method for Assessing the Allergic Contact Dermatitis Potential of Chemicals and Products 05_04A_E1.pdf 05_04A_E2.pdf	OECD TG No. 442B	Jan.2013 JaCVAM statement (LLNA-BraU) First evaluation report on the LLNA:BraU-ELISA for skin sensitization testing Second evaluation report on the LLNA:BraU-ELISA for skin sensitization testing JaCVAM statement (LLNA:BrdU-ELISA) Evaluation report on the Local Lymph Node Assay (LLNA):BraU-ELISA for skin sensitization testing
	05 LLNA:BrdU-FCM	KoCVAM	OECD		OECD TG No. 442B
	06 DPRA	EURL ECVAM Direct Peptide Reactivity Assay (DPRA) ECVAM Validation Study Report Addendum to the Direct Peptide Reactivity Assay (DPRA) ECVAM Validation Study Report	EURL ECVAM		OECD TG No. 442C	Mar.2015 JaCVAM statement Evaluation report on the Direct Peptide Reactivity Assay (DPRA) for alternative to skin sensitization test
	07 ADRA	JaCVAM	JaCVAM			Feb.2024 JaCVAM statement
	08 kDPRA	BASF	EURL ECVAM
	09 KeratinoSense^TM Test Method	EURL ECVAM	EURL ECVAM		OECD TG No. 442D	Aug.2015 JaCVAM statement Evaluation Report on the Alternative Test Method for Skin Sensitization: ARE-Nrf2 Luciferase Test
	10 LuSens^TM Test Method	BASF	EURL ECVAM		OECD TG No. 442D	Nov.2019 JaCVAM statement
	11 h-CLAT Method	JaCVAM & EURL ECVAM	EURL ECVAM		OECD TG No. 442E	Mar.2017 JaCVAM statement Evaluation Report on the Alternative Test Method for Skin Sensitization: human Cell Line Activation Test (h-CLAT)
	12 IL-8 Luc Assay	JaCVAM	JaCVAM			Dec.2020
	13 U-SENS^TM Method	L'Oreal	EURL ECVAM			Nov.2018 JaCVAM statement
	14 GARDskin Test	3RsMC	EURL ECVAM		OECD
	15 SENS-IS^TM Assay	ImmunoSearch			OECD
	16 EpiSensA	JaCVAM	JaCVAM		OECD TG No.422D PS
	17 α-SENS	JaCVAM
	18 Defined Approach for Skin Sensitisation				OECD GL No. 497

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Classification	Test method	Validation study	Peer reviews by experts	Protocol	Test guidelines	Recommendation to government
06 Skin Absorption Test	01 In vitro Skin Absorption Test		OECD Guidance Document for the Conduct of Skin Absorption Studies Basic criteria for the in vitro assessment of dermal absorption of cosmetic ingredients Environmental Health Criteria 235 DERMAL ABSORPTION		OECD TG No. 428 Test No. 428: Skin Absorption: In Vitro Method Basic criteria for the in vitro assessment of dermal absorption of cosmetic ingredients Cosmetic Ingredients: Guidelines for Percutaneous Absorption / Penetration	Jan.2014 JaCVAM statement Evaluation report on the in vitro skin absorption tests

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Classification	Test method	Validation study	Peer reviews by experts	Protocol	Test guidelines	Recommendation to government
07 Acute Oral Toxicity Test	01 Cytotoxicity Test (Estimate Starting Doses for Acute Oral Systemic Toxicity Tests)	EURL ECVAM & ICCVAM	ICCVAM	ICCVAM-Recommended Test Method Protocol BALB/c 3T3 NRU Cytotoxicity Test Method DB-ALM Protocol n° 46 : BALB/c 3T3 Neutral Red Uptake (NRU) Cytotoxicity Test	OECD GD No. 129	Jun.2011 JaCVAM statement Evaluation report on the cytotoxicity tests to estimate starting does for acute oral systemic toxicity tests
	02 Cytotoxicity Test (LD50 ≧ 2000mg/Kg)	EURL ECVAM	EURL ECVAM		EURL ECVAM recommendation	Apr.2019 JaCVAM statement

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Classification	Test method	Validation study	Peer reviews by experts	Test guidelines
08 Genotoxicity Test	01 OECD TG No.473,476, 487,490		OECD	OECD TG No. 473, 476, 487, 490 TG473 TG476 TG487 TG490
	02 In vivo Mammalian Alkaline Comet Assay	JaCVAM Report of the JaCVAM initiative international validation study of the in vivo rodent alkaline Comet assay for the detection of genotoxic carcinogens: the 4th (definitive) phase-1st step Report of the JaCVAM initiative international pre-validation studies of the in vivo rodent alkaline Comet assay for the detection of genotoxic carcinogens	OECD	OECD TG No. 489
	03 Pig-a Assay	US FDA	OECD	OECD TG No. 470
	04 Reconstructed Human Epidermis (RHE) Test Method	Cosmetic Europe/ EURL ECVAM
	05 In vitro Mammalian Alkaline Comet Assay	JaCVAM

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Classification	Test method	Validation study	Peer reviews by experts	Protocol	Test guidelines	Recommendation to government
09 Endcrine Disrupter Screening	01 VM7 Luc ER TA Assay	ICCVAM	ICCVAM	LUMI-CELL® ER ASSAY AGONIST PROTOCOL LUMI-CELL® ER ASSAY ANTAGONIST PROTOCOL DB-ALM Method Summary n° 455: Transactivation Assays to Detect Estrogen Receptor Agonists and Antagonists In Vitro with Stably Transfected Human Cell Lines	OECD TG No. 455	Jan.2014 JaCVAM statement Evaluation report on the BG1Luc estrogen receptor (ER) transactivation (TA) test method (BG1 Luc ER TA)
	02 Stably Transfected Transactivation (STTA) Assay using the (h) ERα-HeLa-9903 cell line	NIHS & CERI	OECD		OECD TG No. 455	Dec.2016 JaCVAM statement Evaluation Report on the Stably Transfected Transcription Activation (STTA) assay using the human Estrogen Receptor (ER)
	03 AR-Ecoscreen	NIHS & CERI 2nd Validation Study Report For Androgen Receptor (AR)-Mediated Stably Transfected Transcriptional Activation (AR-STTA) Assay to Detect Androgenic and Anti-androgenic Activities of Chemicals: AR EcoScreen^TM Draft Report of Pre-validation and Inter-laboratory Validation For Androgen Receptor (AR) Mediated Stably Transfected Transcriptional Activation (AR-STTA) Assay to Detect Androgenic and Anti-androgenic Activities	OECD		OECD TG No. 458	Feb.2019 JaCVAM statement
	04 AR-CALUX	EURL ECVAM	EURL ECVAM
	05 22Rv1/MMTV	Korea	Korea
	06 In vitro Estrogen Receptor Binding Assay	EPA, CERI	OECD		OECD TG No. 493	Dec.2020

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Classification	Test method	Validation study	Peer reviews by experts	Protocol	Test guidelines	Recommendation to government
10 Embryonic Toxicoty Scrneeing	01 Embyonic Stem Cell Test	EURL ECVAM	EURL ECVAM
10 Embryonic Toxicoty Scrneeing	02 Hand1-Luc EST	JaCVAM	JaCVAM		OECD

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Classification	Test method	Validation study	Peer reviews by experts	Protocol	Test guidelines	Recommendation to government
11 Metabolism	01 CYP Induction Method	EURL ECVAM	EURL ECVAM	DB-ALM Protocol n° 193 : Cytochrome P450 (CYP) induction in vitro test method using cryopreserved primary human hepatocytes DB-ALM Protocol n° 194 : Cytochrome P450 (CYP) induction in vitro test method using cryopreserved human HepaRG^TM cell line	OECD
	02 PBK Model				OECD GD No.331

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Classification	Test method	Validation study	Peer reviews by experts	Test guidelines
12 Immunotoxicity Test	01 IL-2 Luc Assay	JaCVAM	JaCVAM	OECD TG No.444A
	02 IL-2 Luc LTT	JaCVAM IL-2 Luc LTT Validation report	JaCVAM IL-2 Luc LTT peer review report
	03 IL-1β Luc assay	JaCVAM

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Classification	Test method	Validation study	Peer reviews by experts	Test guidelines	Recommendation to government
13 Cell Transformation Assay (CTA) for the Screening of Carcinogenitic Activity of Chemicals	01 SHE CTA	EURL ECVAM SYRIAN HAMSTER EMBRYONIC (SHE) CELL PH 7.0 CELL TRANSFORMATION ASSAY PREVALIDATION STUDY REPORT Syrian Hamster Embryonic (SHE) cell pH 7.0 Cell Transformation Assay Prevalidation study Report SYRIAN HAMSTER EMBRYONIC (SHE) CELL PH 6.7 CELL TRANSFORMATION ASSAY PREVALIDATION STUDY REPORT	EURL ECVAM	OECD GD No.214	Mar.2018 Evaluation Report on Syrian Hamster Embryo Cell Transformation Assay
	02 Bhas 42 CTA	EURL ECVAM Bhas 42 Cell Transformation Assay Validation Study Report ECVAM prevalidation study on in vitro cell transformation assays: General outline and conclusions of the study	EURL ECVAM EURL ECVAM Recommendation on the Cell Transformation Assay based on the Bhas 42 cell line ESAC Working Group Peer Review Consensus Report on a Hadano-coordinated validation study of a Cell Transformation Assay (CTA) for carcinogenicity testing based on Bhas 42 cell line	OECD GD No.231	Dec.2020
	03 Balb CTA	EURL ECVAM ECVAM prevalidation study on in vitro cell transformation assays: General outline and conclusions of the study Balb/c 3T3 Cell Transformation Assay Prevalidation study Report	EURL ECVAM	OECD

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Classification	Test method	Validation study	Peer reviews by experts	Test guidelines	Recommendation to government
14 Pyrogen Screening Test	01 PBMC (Peripheral Blood Mononuclear Cell) Assay	EURL ECVAM	EURL ECVAM or ICCVAM ICCVAM Test Method Evaluation Report: Validation Status of Five In Vitro Test Methods Proposed for Assessing Potential Pyrogenicity of Pharmaceuticals and Other Products STATEMENT ON THE VALIDITY OF IN-VITRO PYROGEN TESTS	Ph. Eur.	Evaluation report on in vitro PBMC (peripheral blood mononucleated cell) test method as an alternative to pyrogen testing
	02 PyroMAT法	Merck		Ph.Eur
	03 MylcMAT法	JaCVAM

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Classification	Test method	Validation study	Peer reviews by experts	Protocol	Test guidelines	Recommendation to government
15 Oral Toxicity Test	01 EpiOral^TM