Kinase inhibitor-induced cardiotoxicity assessed in vitro with human pluripotent stem cell derived cardiomyocytes.

Many small molecule kinase inhibitors (SMKIs), used predominantly in cancer therapy, have been implicated in serious clinical cardiac adverse events, which means that traditional preclinical drug development assays were not sufficient for identifying these cardiac liabilities. To improve clinical cardiac safety predictions, the effects of SMKIs targeting many different signaling pathways were studied using human pluripotent stem cell derived cardiomyocytes (hPSC-CMs) in combined assays designed for the detection of both electrophysiological (proarrhythmic) and non-electrophysiological (non-proarrhythmic) drug-induced cardiotoxicity. Several microplate-based assays were used to quantitate cell death, apoptosis, mitochondrial damage, energy depletion, and oxidative stress as mechanism-based non-electrophysiological cardiomyocyte toxicities.

Promoter analysis in ES cell-derived neural cells.

Neural cells derived from ES cells in cell culture (ESNCs) have many of the properties of normal neural cells and provide a model of "neurogenesis-in-a-dish." Here we show that ESNCs provide a powerful system for analyzing neural gene transcription. ES cells are transfected with bacterial artificial chromosomes (BACs) containing Olig2, a gene with a key role in neural fate choice.

Double lox targeting for neural cell transgenesis.

ES cells differentiated along the neural lineage in vitro are an attractive model system. Here we have developed ES cell lines that are suitable for inserting transgenes at a single chromosomal site. ES cell line CE1 (for Cassette Exchange) contains one "acceptor" module (CE1) that allows for efficient double lox targeting.

A subset of ES-cell-derived neural cells marked by gene targeting.

Embryonic stem cells differentiate efficiently in culture into neural progenitors, neurons, oligodendrocytes, and astrocytes. An embryonic stem (ES) cell line with green fluorescent protein (GFP) inserted into the gene for Olig2, a lineage-specific transcription factor, permits visualization and physical separation of a subset of living ES-cell-derived neural cells. GFP-expressing cells have morphological and antigenic properties of the oligodendrocyte lineage.