PEER REVIEW PANEL REPORT:

The Use of In Vitro Basal Cytotoxicity
Test Methods for Estimating Starting Doses for Acute Oral Systemic Toxicity Testing

June 2006

1. Introduction and Rationale for the Use of In Vitro Neutral Red Uptake Cytotoxicity Test Methods to Predict Starting Doses for In Vivo Acute Oral Systemic Toxicity Testing
   
   
   1. Background and Rationale for the Use of In Vitro Cytotoxicity Test Methods to Predict Starting Doses for In Vivo Acute Oral Systemic Toxicity Tests
   2. Regulatory Rationale and Applicability for the Use of In Vitro Cytotoxicity Test Methods to Predict Starting Doses for Acute Oral Systemic Toxicity Testing
   3. Scientific Basis for the In Vitro NRU Test Methods
      
      
2. Test Method Protocol Components of the 3T3 and NHK In Vitro NRU Test Methods
   
   
   1. Overview of the 3T3 and NHK NRU Test Methods
   2. Descriptions and Rationales of the 3T3 and NHK NRU Test Methods
   3. Basis for Selection of the In Vitro NRU Cytotoxicity Test Methods
   4. Proprietary Components of the 3T3 and NHK NRU Test Method Protocols
   5. Basis for Number of Replicate and Repeat Experiments for the 3T3 and NHK NRU Test Methods
   6. Basis for Modifications to the 3T3 and NHK NRU Test Method Protocols
   7. Differences in 3T3 and NHK NRU Test Method Protocols and the Guidance Document Standard Protocols
   8. Overview of the Solubility Protocol
   9. Components of the Solubility Protocol
   10. Basis of the Solubility Protocol
   11. Summary
       
       
3. Reference Substances Used for Validation of the 3T3 and NHK NRU Test Methods
   
   
   1. Rationale for the Reference Substances Selected for Testing
   2. Rationale for the Number of Reference Substances Selected
   3. Characteristics of the Selected Reference Substances
   4. Reference Substance Procurement, Coding, and Distribution
   5. Reference Substances Recommended by the Guidance Document (ICCVAM 2001b)
   6. Summary
      
      
4. In Vivo Rodent Toxicity Reference Values Used to Assess the Accuracy of the 3T3 and NHK NRU Test Methods
   
   
   1. Methods Used to Determine In Vivo Rodent Toxicity Reference Values
   2. Final In Vivo Rodent Toxicity Reference Values
   3. Relevant Toxicity Information for Humans
   4. Accuracy and Reliability of the In Vivo Rodent Toxicity Reference Values
   5. Summary
      
      
5. 3T3 and NHK NRU Test Method Data and Results
   
   
   1. 3T3 and NHK NRU Test Method Protocols
   2. Data Obtained to Evaluate Accuracy and Reliability
   3. Statistical Approaches to the Evaluation of 3T3 and NHK NRU Data
   4. Summary of Results
   5. Coded Reference Substances and GLP Guidelines
   6. Study Timeline and NICEATM/ECVAM Study Participatory Laboratories
   7. Availability of Data
   8. Solubility Test Results
   9. Summary
      
      
6. Accuracy of the 3T3 and NHK NRU Test Methods
   
   
   1. Accuracy of the 3T3 and NHK NRU Test Methods for Predicting Acute Oral Systemic Toxicity
   2. Improving the Prediction of Rodent LD50 Values from In Vitro NRU IC50 Data
   3. Accuracy of the 3T3 and NHK NRU Test Methods for Toxicity Category Predictions
   4. Strengths and Limitations of the 3T3 and NHK NRU Test Methods for In Vivo Toxicity Prediction
   5. Salient Issues of Data Interpretation
   6. Comparison to Established Performance Standards
   7. Summary
      
      
7. Reliability of the 3T3 and NHK NRU Test Methods
   
   
   1. Substances Used to Determine the Reliability of the 3T3 and NHK NRU Test Methods
   2. Reproducibility Analyses for the 3T3 and NHK NRU Test Methods
   3. Historical Positive Control Data
   4. Laboratory Concordance for Solvent Selection
   5. Summary
      
      
8. 3T3 and NHK NRU Test Method Data Quality
   
   
   1. Adherence to Good Laboratory Practice Guidelines
   2. Results of Data Quality Audits
   3. Impact of Deviations from GLPs/Non-compliance
   4. Availability of Laboratory Notebooks
   5. Summary
      
      
9. Other Scientific Reports and Reviews of In Vitro Cytotoxicity Test Methods and the Ability of These Test Methods to Predict Acute Systemic Toxicity
   
   
   1. Relevant Studies
   2. Independent Scientific Reviews
   3. Studies Using In Vitro Cytotoxicity Test Methods with Established Performance Standards
   4. Summary
      
      
10. Animal Welfare Considerations (Refinement, Reduction, and Replacement)
    
    
    1. Use of 3T3 and NHK NRU Test Methods to Predict Starting Doses for Acute Systemic Toxicity Assays
    2. Reduction and Refinement of Animal Use for the UDP
    3. Reduction and Refinement of Animal Use for the ATC
    4. Summary
       
       
11. Practical Considerations
    
    
    1. Transferability of the 3T3 and NHK NRU Test Methods
    2. 3T3 and NHK NRU Test Method Training Considerations
    3. Test Method Cost Considerations
    4. Time Considerations for the 3T3 and NHK NRU Test Methods
    5. Summary