Misiuta I E, Saporta S, Sanberg P R, Zigova T, Willing A E
Center of Excellence for Aging and Brain Repair, University of South Florida, Tampa, Florida, USA.
J Neurosci Res. 2006 Mar;83(4):668-79. doi: 10.1002/jnr.20718.
Our laboratory is working with the human NTera2/D1 (NT2) cell line, which has properties similar to those of progenitor cells in the central nervous system (CNS). These neural-like precursor cells can differentiate into all three major lineages, neurons, astrocytes, and oligodendrocytes. The pure neuronal population, hNT neurons, possess characteristics of dopamine (DA) cells. First, we analyzed whether the retinoic acid (RA)-treated hNT neurons and the NT2 precursor cells expressed two transcription factors required for development of the midbrain DA neurons. We report that NT2 cells endogenously expressed Engrailed-1 and Ptx3, whereas RA-treated hNT neurons did not express Engrailed-1 or Ptx3. Next we examined the influence of lithium treatment on Engrailed-1 and Ptx3 as well as another critical transcription factor, Nurr1. Previous research has shown that lithium can mimic the Wnt pathway, which is important for the induction of these transcription factors. Finally, we investigated the effect of lithium treatment on the viability and proliferation of NT2 cells, because lithium has been shown to stimulate neurogenesis in adult neural precursors. Lithium treatment increased the viability and proliferation of NT2 cells. The expression of transcription factors essential for the induction and maintenance of the DA phenotype was not increased in NT2 after lithium treatment. We conclude that the NT2 cell line is an excellent in vitro model system for studying the influence of pharmalogical agents on proliferation, differentiation, and apoptosis of a human neural progenitor cell line.
我们的实验室正在研究人NTera2/D1（NT2）细胞系，该细胞系具有与中枢神经系统（CNS）中祖细胞相似的特性。这些神经样前体细胞可分化为所有三种主要谱系，即神经元、星形胶质细胞和少突胶质细胞。纯神经元群体，即hNT神经元，具有多巴胺（DA）细胞的特征。首先，我们分析了经视黄酸（RA）处理的hNT神经元和NT2前体细胞是否表达中脑DA神经元发育所需的两种转录因子。我们报告称，NT2细胞内源性表达Engrailed-1和Ptx3，而经RA处理的hNT神经元不表达Engrailed-1或Ptx3。接下来，我们研究了锂处理对Engrailed-1、Ptx3以及另一个关键转录因子Nurr1的影响。先前的研究表明，锂可模拟Wnt信号通路，而该信号通路对这些转录因子的诱导很重要。最后，我们研究了锂处理对NT2细胞活力和增殖的影响，因为锂已被证明可刺激成年神经前体细胞的神经发生。锂处理增加了NT2细胞的活力和增殖。锂处理后，NT2中诱导和维持DA表型所需的转录因子表达并未增加。我们得出结论，NT2细胞系是研究药物对人神经祖细胞系的增殖、分化和凋亡影响的优秀体外模型系统。
