Appendix C Table of Contents 1.0 STANDARD OPERATING PROCEDURE (SOP) FOR THE BALB/C 3T3 NEUTRAL RED UPTAKE CYTOTOXICITY TEST - A TEST FOR BASAL
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چکیده
1.0 STANDARD OPERATING PROCEDURE (SOP) FOR THE BALB/C 3T3 NEUTRAL RED UPTAKE CYTOTOXICITY TEST A TEST FOR BASAL CYTOTOXICITY....................... 3 1.1 Background and Introduction.................................................................................................. 3 1.2 Rationale..................................................................................................................................... 3 1.3 Basic Procedure ......................................................................................................................... 3 1.4 Test Limitations ......................................................................................................................... 4 1.5 Material ...................................................................................................................................... 4 1.5.1 Cell Lines ............................................................................................................................ 4 1.5.2 Technical Equipment .......................................................................................................... 4 1.5.3 Chemicals, Media, and Sera ............................................................................................... 4 1.5.4 Preparations ....................................................................................................................... 5 1.5.4.1 Media............................................................................................................................ 5 1.5.4.2 Neutral Red (NR) Stock Solution.................................................................................. 5 1.5.4.3 Neutral Red (NR) Medium............................................................................................ 5 1.5.4.4 Ethanol/Acetic Acid Solution (NR Desorb) .................................................................. 5 1.5.4.5 Preparation of Test Chemicals..................................................................................... 6 1.6 Methods ...................................................................................................................................... 6 1.6.1 Cell Maintenance and Culture Procedures ........................................................................ 6 1.6.1.1 Routine Culture of BALB/C 3T3 Cells ......................................................................... 6 1.6.1.2 Cell Counting ............................................................................................................... 6 1.6.1.3 Subculture..................................................................................................................... 6 1.6.1.4 Freezing........................................................................................................................ 7 1.6.1.5 Thawing ........................................................................................................................ 7 1.6.2 Quality Check of Assay (I): Positive Control (PC)............................................................. 7 1.6.3 Quality Check of Assay (II): Vehicle Control (VC) ............................................................ 7 1.6.4 Quality Check of Concentration-Response ......................................................................... 8 1.6.5 Concentrations of Test Chemical........................................................................................ 8 1.6.5.1 Range Finder Experiment ............................................................................................ 8 1.6.5.2 Main Experiment .......................................................................................................... 8 1.6.6 Test Procedure .................................................................................................................... 8 1.7 Data Analysis............................................................................................................................ 10 1.8 Prediction Model...................................................................................................................... 11 1.9 References................................................................................................................................. 11
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