Monitoring neuronal differentiation in the embryonal carcinoma cell line NTERA2

Abstract

NTERA2 embryonal carcinoma cells are pluripotent cells that originated from a metastasis of a testicular cancer. Previous research has demonstrated the ability of NTERA2 cells to reliably differentiate into a neuronal cell fate, upon exposure to retinoic acid. Although this provides a useful tool for the study of neuronal differentiation, until now a thorough investigation of basic in vitro conditions was absent. This study has systematically analysed NTERA2 differentiation as a population and at the single cell level. Prolonged RA exposure and high cell density has been shown to be required for the successful production of neurons, demonstrating that NTERA2 cells follow a nonautonomous path of differentiation towards a neuronal phenotype. Gap junction communication is required for NTERA2 neuronal differentiation and is thought to contribute to the density dependence of neuronal differentiation. The importance of Wnt signalling has also been examined and it has been found that elevated Wnt signalling drives RA mediated cell differentiation to a non-neuronal fate. Single cell analysis has revealed that functional heterogeneity resides within the undifferentiated stem cell population. SSEA3 expression was successfully used to enrich for functional stem cells however examination of SSEA3 expression does not enable the prediction of single cell behaviour. Diversity and instability of chromosomal karyotype in the NTERA2 cell line suggests that single cell heterogeneity is partially derived from karyotypic instability.EThOS - Electronic Theses Online ServiceGBUnited Kingdo