title: Characterisation of neurons derived from a cortical human neural stem cell line CTX0E16.
abstract: BACKGROUND
  Conditionally immortalised human neural progenitor cells (hNPCs) represent a robust source of native neural cells to investigate physiological mechanisms in both health and disease. However, in order to recognise the utility of such cells, it is critical to determine whether they retain characteristics of their tissue of origin and generate appropriate neural cell types upon differentiation. To this end, we have characterised the conditionally immortalised, cortically-derived, human NPC line, CTX0E16, investigating the molecular and cellular phenotype of differentiated neurons to determine whether they possess characteristics of cortical glutamatergic neurons.
  
  
  METHODS
  Differentiated CTX0E16 cells were characterised by assessing expression of several neural fates markers, and examination of developing neuronal morphology. Expression of neurotransmitter receptors, signalling proteins and related proteins were assessed by q- and RT-PCR and complemented by Ca(2+) imaging, electrophysiology and assessment of ERK signalling in response to neurotransmitter ligand application. Finally, differentiated neurons were assessed for their ability to form putative synapses and to respond to activity-dependent stimulation.
  
  
  RESULTS
  Differentiation of CTX0E16 hNPCs predominately resulted in the generation of neurons expressing markers of cortical and glutamatergic (excitatory) fate, and with a typical polarized neuronal morphology. Gene expression analysis confirmed an upregulation in the expression of cortical, glutamatergic and signalling proteins following differentiation. CTX0E16 neurons demonstrated Ca(2+) and ERK1/2 responses following exogenous neurotransmitter application, and after 6 weeks displayed spontaneous Ca(2+) transients and electrophysiological properties consistent with that of immature neurons. Differentiated CTX0E16 neurons also expressed a range of pre- and post-synaptic proteins that co-localized along distal dendrites, and moreover, displayed structural plasticity in response to modulation of neuronal activity.
  
  
  CONCLUSIONS
  Taken together, these findings demonstrate that the CTX0E16 hNPC line is a robust source of cortical neurons, which display functional properties consistent with a glutamatergic phenotype. Thus CTX0E16 neurons can be used to study cortical cell function, and furthermore, as these neurons express a range of disease-associated genes, they represent an ideal platform with which to investigate neurodevelopmental mechanisms in native human cells in health and disease.
authors: Department of Basic and Clinical Neuroscience, Institute of Psychiatry Psychology & Neuroscience, King's College London, London, SE5 8AF, UK. greg.anderson@kcl.ac.uk.
authors: Greg W
authors: GW
authors: Anderson
authors: Department of Basic and Clinical Neuroscience, Institute of Psychiatry Psychology & Neuroscience, King's College London, London, SE5 8AF, UK. peter.deans@kcl.ac.uk.
authors: P J Michael
authors: PJ
authors: Deans
authors: MRC Centre for Developmental Neurobiology, King's College London, London, SE5 8AF, UK. ruth.taylor@kcl.ac.uk.
authors: Ruth D T
authors: RD
authors: Taylor
authors: Department of Basic and Clinical Neuroscience, Institute of Psychiatry Psychology & Neuroscience, King's College London, London, SE5 8AF, UK. pooja.1.raval@kcl.ac.uk.
authors: Pooja
authors: P
authors: Raval
authors: Department of Basic and Clinical Neuroscience, Institute of Psychiatry Psychology & Neuroscience, King's College London, London, SE5 8AF, UK. dchenjx@gmail.com.
authors: Ding
authors: D
authors: Chen
authors: Department of Basic and Clinical Neuroscience, Institute of Psychiatry Psychology & Neuroscience, King's College London, London, SE5 8AF, UK. lowderhb@gmail.com.
authors: Harrison
authors: H
authors: Lowder
authors: Department of Basic and Clinical Neuroscience, Institute of Psychiatry Psychology & Neuroscience, King's College London, London, SE5 8AF, UK. srishtimukherji@yahoo.co.uk.
authors: Srishti
authors: S
authors: Murkerji
authors: MRC Centre for Developmental Neurobiology, King's College London, London, SE5 8AF, UK. laura.andreae@kcl.ac.uk.
authors: Laura C
authors: LC
authors: Andreae
authors: Department of Basic and Clinical Neuroscience, Institute of Psychiatry Psychology & Neuroscience, King's College London, London, SE5 8AF, UK. brenda.p.williams@kcl.ac.uk.
authors: Brenda P
authors: BP
authors: Williams
authors: Department of Basic and Clinical Neuroscience, Institute of Psychiatry Psychology & Neuroscience, King's College London, London, SE5 8AF, UK. deepak.srivsatava@kcl.ac.uk.
authors: Deepak P
authors: DP
authors: Srivastava
mesh_terms: D000200:Action Potentials / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D006801:Humans; D058953:Neural Stem Cells / Q000166:cytology*; D009474:Neurons / Q000166:cytology* / Q000378:metabolism