Sullivan Kathryn M, Park Chang Gyun, Ito John D, Kandel Mikhail, Popescu Gabriel, Kim Young Jun, Kong Hyunjoon
Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, United States.
Institute of Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, United States.
Environ Sci Technol. 2020 Nov 3;54(21):13797-13806. doi: 10.1021/acs.est.0c01988. Epub 2020 Oct 19.
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.
内分泌干扰化学物质（EDC）包括模拟天然激素结构或功能的合成化合物。虽然大多数研究使用活胚胎或成年鱼的原代细胞，但当这些细胞在涂有细胞外基质蛋白的塑料或玻璃基质上培养时，它们会迅速失去功能。本研究假设，粘附鱼细胞的基质的柔软度可以在EDC暴露期间调节工程化肝样组织的细胞间组织和生理功能。我们通过在具有可控弹性模量的基于胶原蛋白的水凝胶上培养斑马鱼肝细胞（ZF-L），通过检查暴露于17β-雌二醇的肝样组织的形态、尿素产生和细胞内氧化应激，仔细研究了这一假设。有趣的是，与较硬的凝胶相比，较软的凝胶促使细胞形成具有类似胆小管结构的细胞片。在较软凝胶上培养的肝样组织在暴露于17β-雌二醇时表现出更活跃的尿素产生，并通过梯度光干涉显微镜（GLIM，一种活细胞成像技术）证实细胞内活性氧水平恢复更快。这些结果对于改进对潜在EDC对水生生物和人类健康影响的筛查和理解具有广泛的用途。
