Characterization of the Dopaminergic Potential of the Human NTera2/D1 (NT2) Cell Line <em>In Vitro</em>

• Main Author: Misiuta, Iwona E
• Format: Others
• Published: Scholar Commons 2005
• Subjects: hNT; NT2; dopamine; development; Parkinson’s disease; Engrailed; Ptx3; Nurr1; lithium; survival; proliferation; differentiation; neurite outgrowth; American Studies; Arts and Humanities
• Online Access: https://scholarcommons.usf.edu/etd/775; https://scholarcommons.usf.edu/cgi/viewcontent.cgi?article=1774&amp;context=etd

Our laboratory is working with the human NTera2/D1 (NT2) cell line which have properties similar to progenitor cells in the CNS. These neural-like precursors cells can differentiate into all three major lineages - neurons, astrocytes, andoligodendrocytes. The pure neuronal population, called the hNT cells, possess characteristics of dopamine (DA) cells. In this dissertation, we performed various experiments to examine the neuronal and dopaminergic development of this cellline. We first cultured our hNT neurons with cells from the developingnigrostriatal (NS) pathway, the ventral mesencephalon and striatum, to determine their influence on survival, neuritic outgrowth, and DA phenotype. The survival ofhNT neurons was substantially greater when they were cultured with embryonicday (E) 18 cells, compared to monocultures or cocultures with either E14 orpostnatal day (P) 1 cells. The neuritic outgrowth of hNT neurons as assessed by the number of primary neurites per cell was increased when cultured with theareas of the brain from E14 and P1. The DA phenotype, as determined by the expression of the rate-limiting enzyme of DA synthesis was not increased in hNTneurons when they were cultured with primary rat cells from the NS pathway.Next we analyzed if the retinoic acid (RA)-treated hNT neurons and the NT2 precursor cells expressed three transcription factors required for development ofthe DA phenotype. We report that NT2 cells endogenously expressed Engrailed-1, Ptx3, and Nurr1 while RA treatment increased Nurr1 but down-regulated Engrailed-1 and Ptx3. Finally, lithium has been shown to stimulate neurogenesisin adult hippocampal precursors as well as influence the Wnt pathway known to be important for the induction of the DA phenotype.