Figure 5. Sentinel cell design to ascertain batch-to-batch responsiveness, assay performance and to eliminate risk of failure during HTS.

A. HTS workflow. From a single vial of primary cells, a small portion is set aside after thawing and left unstimulated or stimulated for 3 days with DOX to induce expression of lamin B1. The majority of cells is expanded into multiple flasks and stimulated for experiments after the second split. Because the sentinel cells are from the same vial and are processed before the large batch gets exposed to compounds, unresponsive batches are identified before a large scale screen is executed, eliminating the risk of losing precious library compounds during HTS. B. Assessment of batch-to-batch variability. Table shows canonical HTS measures obtained on different days with different batches of TRE-MEF. Data for each experiments are based on four technical replicates.