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A Cell Model to Evaluate Chemical Effects on Adult Human Cardiac Progenitor Cell Differentiation and Function
Das, K., V. Long III, M. Kubo, C. Lau, AND K. Dreher. A Cell Model to Evaluate Chemical Effects on Adult Human Cardiac Progenitor Cell Differentiation and Function. 57th SOT Annual Meeting, San Antonio, Texas, March 11 - 15, 2018.
The main purpose of this study to establish an alternative testing model to evaluate the effect of chemicals on adult human CPC differentiation to CM and their function. Study provides information related to the impact of chemicals and environmental exposure on human longevity.
Adult cardiac stem cells (CSC) and progenitor cells (CPC) represent a population of cells in the heart critical for its regeneration and function over a lifetime. The impact of chemicals on adult human CSC/CPC differentiation and function is unknown. Research was conducted to develop an improved assay to assess the impact of chemicals on CPC differentiation and function. CPCs (Cellular Dynamics International Inc.) were treated with 10 µM XAV939(Wnt/β-catenin inhibitor) + 2.5 µM SB431542 (TGF-βR inhibitor) for 48h. Cultures were then maintained in either Williams Eagle (WE) or maturation medium (MM) with or without T3 for 2 - 21 days to identify the optimal conditions for CPC differentiation, cardiomyocytes (CM) function, and compare to manufacturer’s recommended protocol. CPC cytotoxicity and differentiation were determined by quantitative fluorescent analysis using Red Dot1 nuclear and anti-cardiac troponin T antibody (TnT) in-cell western staining (LI-COR Odyssey CLx Imaging System), respectively. CM function was determined by measuring beat rate (BR) and beat amplitude (BAMP) using the xCELLigence RTCA Cardio platform (ACEA Biosciences). Impacts of environmental chemicals on CPC differentiation and CM function were evaluated in these optimized conditions. Cell viability was not affected at any time by any media condition tested. Maximal CPC differentiation measured by TnT expression occurred in MM medium after 8 days and represented a 74% increase of TnT expression versus the manufacturer’s protocol. Maximal CM function was detected in MM after 21 days with a 29% and 50% increase in BR and BAMP, respectively, versus the manufacturer’s protocol. CPCs exposed to perfluorononanoate (25, 50, 100 µM), Triclosan (2.5, 5, 10 µM), or Bisphenol A (0.125, 1.25, 12.5 µM) using optimized CPC assay conditions led to a 25% decrease in BR and BAMP at 13 and 21 days post-exposure, at the lowest concentration tested. This research provides a novel alternative testing model to evaluate the effect of chemicals on adult human CPC differentiation to CM and their function. Our results demonstrate that chemicals significantly impact the ability of CPC to differentiate into functional CM. This adverse effect may impact the heart in terms of susceptibility to disease and longevity. (This abstract does not represent EPA policy)