High Content Screening to Study Cellular Response to Treatment by Using Primary In Vitro Cultures Established from CNS Tumour Samples

Abstract

This study investigated if in vitro cultures of cancer stem cells derived from paediatric high-grade glioma tumours can be used to study the efficiency of therapeutic drugs by using custom analysis sequences with the Harmony software developed by Operetta. Four different drugs - vinorelbine, vincristine, etoposide, and methotrexate –, with and without irradiation, were tested. The nuclei were divided into subpopulations based on morphological and intensity properties with the hypothesis that each subpopulation corresponded to a specific cell state. The study also tested the theory that a subpopulation composed of presumably dead cells could be identified and used to discern drug effects not detected by studying the viable cell population alone. We found that each subpopulation demonstrated unique responses to the different treatments in concurrence with different cell states, showing that this method of subtyping has potential uses. It also demonstrates the possibility to use the subpopulation of dead cells in order to detect drug efficacy not readily apparent from observation of the total number of cells.