Matrix Softness-Mediated 3D Zebrafish Hepatocyte Modulates Response to Endocrine Disrupting Chemicals

Abstract

Endocrine disrupting chemicals (EDC) include synthetic compounds that mimic the structure or function of natural hormones. While most studies utilize live embryos or primary cells from adult fish, these cells rapidly lose functionality when cultured on plastic or glass substrates coated with extracellular matrix proteins. This study hypothesizes that the softness of a matrix with adhered fish cells can regulate the intercellular organization and physiological function of engineered hepatoids during EDC exposure. We scrutinized this hypothesis by culturing zebrafish hepatocytes (ZF-L) on collagen-based hydrogels with controlled elastic moduli by examining morphology, urea production, and intracellular oxidative stress of hepatoids exposed to 17β-estradiol. Interestingly, the softer gel drove cells to form a cell sheet with a canaliculi-like structure compared to its stiffer gel counterpart. The hepatoids cultured on the softer gel exhibited more active urea production upon exposure to 17β-estradiol and displayed faster recovery of intracellular reactive oxygen species level confirmed by gradient light interference microscopy (GLIM), a live-cell imaging technique. These results are broadly useful to improve screening and understanding of potential EDC impacts on aquatic organisms and human health.

Multiscale Assay of Unlabeled Neurite Dynamics Using Phase Imaging with Computational Specificity

“…To acquire the quantitative phase images, we use gradient light interference microscopy (GLIM), which measures the optical path length shifts associated with the specimen in a differential interference contrast (DIC) geometry. In short, GLIM was used in this study to measure quantitative parameters, improving the image quality in DIC while maintaining the fluorescence signal . Furthermore, the GLIM module is a relatively straightforward modification to the conventional DIC microscope (Supplementary Note 1).…”

“…In short, GLIM was used in this study to measure quantitative parameters, improving the image quality in DIC while maintaining the fluorescence signal. 42 Furthermore, the GLIM module 43 is a relatively straightforward modification to the conventional DIC microscope (Supplementary Note 1).…”