HTS/HCS to Screen Molecules Able to Maintain Embryonic Stem Cell Self-Renewal or to Induce Differentiation: Overview of Protocols

Abstract

Embryonic stem (ES) cells, combining self-renewal ability with wide range tissue-specific cell differentiation, represent one of the most powerful model systems in basic research, drug discovery and biomedical applications. In the field of drug development, ES cells are instrumental in high-throughput/content screening (HTS/HCS) for the evaluation of large compound libraries to test biological activity and toxic properties. Since it is a high priority to test new compounds in vitro, before starting animal and human treatments, there is an increasing demand for new in vitro models that can be used in HTS/HCS to facilitate drug development. In order to achieve this objective, several methods for ES cell self-renewal or differentiation have been evaluated to assess their compatibility with HTS/HCS. This review describes protocols used to screen molecules able to maintain self-renewal or to induce differentiation in ectodermal, mesodermal, endodermal, and their derivative cell lines.

A Chemical Biology Study of Human Pluripotent Stem Cells Unveils HSPA8 as a Key Regulator of Pluripotency

“…The earliest attempts of pluripotency regulator screenings adopted mouse embryonic stem cells (mESCs) and mouse embryonal carcinoma cells (mECCs) as their screening platforms ( Chen et al., 2006 , Ding et al., 2003 , Zhu et al., 2009 ). More recently, several hESC- and hiPSC-based HTSs have been reported ( Barbaric et al., 2010 , Ben-David et al., 2013 , Desbordes et al., 2008 , Gonzalez et al., 2011b , Kameoka et al., 2014 , Kumagai et al., 2013 , Manganelli et al., 2014 , Xu et al., 2010 ). Compared with those platforms, we believe that hECCs have been underappreciated for HTS.…”

“…In recent years, hESCs and human induced pluripotent stem cells (hiPSCs) have been used successfully for HTS in several studies ( Barbaric et al., 2010 , Ben-David et al., 2013 , Desbordes et al., 2008 , Gonzalez et al., 2011b , Kameoka et al., 2014 , Kumagai et al., 2013 , Manganelli et al., 2014 , Xu et al., 2010 ). However, the high cost associated with the maintenance and scale-up of human pluripotent stem cells (hPSCs) inevitably limits the scale of their application in HTS studies.…”