A Tph2GFP Reporter Stem Cell Line To Model in Vitro and in Vivo Serotonergic Neuron Development and Function.
Giulia PaciniAttilio MarinoSara MigliariniElisa BrilliBarbara PelosiGiacomo MaddaloniMarta PratelliMario PellegrinoAldo FerrariMassimo PasqualettiPublished in: ACS chemical neuroscience (2017)
Modeling biological systems in vitro has contributed to clarification of complex mechanisms in simplified and controlled experimental conditions. Mouse embryonic stem (mES) cells can be successfully differentiated toward specific neuronal cell fates, thus representing an attractive tool to dissect, in vitro, mechanisms that underlie complex neuronal features. In this study, we generated and characterized a reporter mES cell line, called Tph2GFP, in which the vital reporter GFP replaces the tryptophan hydroxylase 2 (Tph2) gene. Tph2GFP mES cells selectively express GFP upon in vitro differentiation toward the serotonergic fate, they synthesize serotonin, possess excitable membranes, and show the typical morphological, morphometrical, and molecular features of in vivo serotonergic neurons. Thanks to the vital reporter GFP, we highlighted by time-lapse video microscopy several dynamic processes such as cell migration and axonal outgrowth in living cultures. Finally, we demonstrated that predifferentiated Tph2GFP cells are able to terminally differentiate, integrate, and innervate the host brain when grafted in vivo. On the whole, the present study introduces the Tph2GFP mES cell line as a useful tool allowing accurate developmental and dynamic studies and representing a reliable platform for the study of serotonergic neurons in health and disease.
Keyphrases
- mesenchymal stem cells
- induced apoptosis
- crispr cas
- cell cycle arrest
- cell migration
- public health
- high resolution
- mental health
- single molecule
- spinal cord injury
- gene expression
- signaling pathway
- dna methylation
- cell death
- mass spectrometry
- oxidative stress
- cell proliferation
- risk assessment
- multiple sclerosis
- cerebral ischemia
- health information