Apoptosis | InSphero

Mechanistic Assays


Utilize Caspase 3/7 activation as a sensitive marker for hepatotoxicity

Human Liver Spheroid model for toxicology testing

Drug-induced apoptosis of hepatocytes is one of the earliest hallmarks of liver injury. Apoptosis is the first step towards hepatocellular death, and plays a key role in hepatic inflammation and fibrogenesis (1). Unlike plated 2D hepatocytes, which are at least partially resistant to apoptosis due to activation of PI3K/Akt pathway (1), 3D InSight™ Human Liver Microtissues maintain susceptibility towards induction of apoptosis, making it possible to detect early signs of hepatotoxicity with sensitive cell based-assays.

  • Retain sensitivity to apoptosis with primary human hepatocytes
  • Detect early apoptotic events that precede the drop of intracellular ATP-content or release of enzyme leakage markers
  • Use sensitive assay technology (Caspase 3/7-Glo®, Promega) to evaluate drug-induced apoptosis in a throughput compatible, cost-effective manner


  1. Godoy et al, Arch Toxicol (2013)

Example Data

Caspase 3/7 activation is an early marker of hepatocyte cell death. Microtissues were treated with Aflatoxin B1 for 24 hours, and intracellular ATP levels, intracellular Caspase 3/7 activity and alpha- GST release were evaluated.  ATP levels are shown as % of control, while levels of Caspase 3/7 and a-GST are shown in arbitrary units (AU). Activation of Caspase 3/7 preceded the decline in ATP-content and alpha-GST release into the supernatant, suggesting that caspase 3/7 is a sensitive marker for drug-induced apoptosis.

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