TACS MTT Cell Proliferation Assay

MTT Assay of Human Tissue-derived Liver Organoids. Human Liver organoids were derived from human biopsy tissue. Undifferentiated liver organoids were formed by embedding dissociated tissue in Cultrex RGF BME, Type 2 (Catalog # 3533-005-02) and culturing in specialized media. Liver organoids were differentiated using media containing Recombinant Human FGF-19 (Catalog # 969-FG), DAPT (Catalog # 2634), and Dexamethasone (Catalog # 1126). A) Representative images of untreated control and DMSO-treated (positive control) undifferentiated and differentiated liver organoids. B) Quantification of MTT staining of cells treated with varying doses of acetaminophen, Trovafloxin, or Doxorubicin showed a dose-dependent loss of cell viability under each condition. Interestingly, liver organoids showed increased vulnerability to hepatotoxicity as they matured in differentiation media.

Toxicity analysis of iPSC-derived Hepatocytes using TACS MTT Proliferation Assay and the CometChip Assay. iPSC-derived hepatocytes were generated using the StemXVivo Hepatocyte Differentiation Kit (Catalog # SC033). iPSC-derived hepatocytes were treated with Acetaminophen, Trovafloxacin, and Doxorubicin and dose response toxicity profiles using the (A) TACS MTT Cell Proliferation Assay or (B) CometChip (Catalog # 4260-096-ESK). Using the MTT assay, Trovafloxacin and Doxorubicin showed significantly increased loss of cell metabolic activity at 100 µM and >10 µM, respectively. Increased DNA damage was detected at these two concentrations using the CometChip (n ≥ 3 for each group).

HepG2 Spheroid Toxicity Detected using the MTT Assay. A) Hepatocyte spheroids were formed by plating HepG2 liver hepatocellular carcinoma cells (10,000 cells per well) in a 24-well plate coated with Cultrex RGF BME, Type 2 (Catalog # 3533-005-02). B) HepG2 spheroids were cultured for 21 days prior exposure to acetaminophen, trovafloxacin, or doxorubicin. Spheroids showed significantly increased loss of cell metabolic activity at 100 µM and >10 µM, respectively (n ≥ 3 for each group), as detected by the MTT Assay.