Technical evaluation and standardization of the human thyroid microtissue assay

The success and sustainability of U.S. EPA efforts to reduce, refine, and replace in vivo animal testing depends on the ability to translate toxicokinetic and toxicodynamic data from in vitro and in silico new approach methods (NAMs) to human-relevant exposures and health outcomes. Organotypic culture models employing primary human cells enable consideration of human health effects and inter-individual variability, but present significant challenges for test method standardization, transferability, and validation. Increasing confidence in the information provided by these in vitro NAMs requires setting appropriate performance standards and benchmarks, defined by the context of use, to consider human biology and mechanistic relevance without animal data. The human thyroid microtissue assay utilizes primary human thyrocytes to reproduce structural and functional features of the thyroid gland that enable testing for potential thyroid disrupting chemicals. As a variable-donor assay platform..., , , # Technical Evaluation and Standardization of the Human Thyroid Microtissue Assay

**SUPPLEMENTARY TABLES LEGENDS**
 
**Supplementary Table 1. Human Thyroid Microtissue (hTMT) Medium.** Formulation of hTMT medium components, vendor, catalog number, and final working concentrations.   
 
**Supplementary Table 2. Assay Performance Parameters.** Performance criteria were established from the T4 median, median absolute deviation (MAD), and range values derived from the qualified donor cohort under assay-specific conditions with required priority levels for all metrics. 
 
**Supplementary Table 3. Donor Procurement Parameters.** Donor procurement specifications extrapolated from linear regression modeling using the acceptance criteria defined for thyroglobulin, thyroxine (T4), and triiodothyronine (T3). Not applicable (NA).
 
**Supplementary Table 4. Multiple Linear Regression Statistics.** Multiple linear regression model statistics for microtissue biomass, thyroglobulin, T4, T3, and T4/T3 ...