1. ASTM Subcommittee on Cell Signaling (F04.46) Established May 2006 Anne Plant, NIST Norfolk VA May 24, 2007

2. TE products depend on achieving desired response of cells Cell response is complicated: many parameters, largely undefined Cells are extremely sensitive to their environment (ECM, scaffold, in vivo conditions) WHY Standards for Cell Signaling? Copyright © 2000 Cell Press. The Hallmarks of Cancer Douglas Hanahan and Robert A. Weinberg

3. Purpose of the Subcommittee on Cell Signaling Directing cells to migrate, differentiate, and assemble in a desired fashion may require understanding of the intracellular pathways that control the response of cells to a complex environment. Accurate and quantitative measurements of cell signaling biomarkers, aided by reference materials and standards would aid R&D (development of hypotheses, prediction of outcome, comparison of data over time). Would aid regulatory, and QA/QC (by providing predictive capabilities for determining safety, efficacy). More than one biomarker may be needed to evaluate a response. Standardized assays would facilitate pooling of data.

4. Stages of product development R&D: Must have quantitative methods for comparing cells day to day to evaluate differentiation, response to conditions and materials QA/QC: Must have quantitative methods to evaluate health of cells, predict future response, determine consistency WHY Standards for Cell Signaling?

5. : Classification of Signaling Pathways Ex: kinases, integrin-dependent, G-protein coupled, mechanical response, differentiation, etc. Cell Signaling Mechanism-Based assays for Cytotoxicity Greater predictive capability compared to live/dead Terminology and Definitions (Horowitz) Standard Protocols for Extracellular Matrix Preparation and Presentation Expected response from cells under controlled conditions Stable Storage of Cells Integrity wrt genetics, response to environment, ‘health’. Cell Identification markers Standard conditions, cell normalization, defined media Quantitative methods for comparison of cells (Elliott) Task areas previously under consideration: Some of these topics may be well-developed, some may be research areas. Comments? Topics in RED were added during Atlanta meeting spring 2006

6. Cell morphology and optical ‘mask’ for quantifying intracellular markers [Elliott] Statistical analysis for cell population distributions Quantification of GFP in reporter cells Cell volume as an indicator of growth rate, division time Underpinning protocols: Quantitative methods for comparison of cells

7. Goals for this Subcommittee meeting 1.Report on draft guidance document on protocol for measuring morphology John Elliott, NIST 2.Report on terminology Manny Horowitz, Johns Hopkins University 3.Populate task groups

8. Classification of Signaling Pathways (and associated informatics) Cell Signaling Mechanism-Based assays for Cytotoxicity Stress (Moos, Kaplan, Durfor, Doser, Williams) Terminology and Definitions (Horowitz) Standard Protocols for Extracellular Matrix Preparation and Presentation for controlling signaling (Lee, Durfor, Kaplan, Elliott) Cell Identification markers (Moos) Stable Storage of Cells (Suh) Standard conditions, defined media (Suh) Quantitative methods for comparison of cells (Elliott) Criteria for biomarker validation (measurement, clinical relevance, multiple markers) (McFarland, Moos, Elliott) Matrix remodeling Cell Proliferation Updated Task Areas: Standards, protocols or guidance for… Updated Norfolk, 5/24/07. Changes to previous list are in red (added) or grey (deleted). Task force contributors are in parentheses.

9. Other discussion topics that arose: Using documents from other organizations such as AATB; and encouraging individuals from such organizations to participate in ASTM (Suh) [Additional input welcome] Tumorogenicity (this might be incorporated into the ‘Stress’ topic) (Durfor) Symposium on Cell Signaling Measurements (follow-up to 2003 symposium). To be held at NIST? Sometime in 2008? (Horowitz, Elliott) [Additional input welcome] Your input is critical! All task areas are open to anyone who is interested in participating. Next meeting is during ASTM meeting week November 13-16, Tampa FL