中国神经再生研究(英文版) ›› 2026, Vol. 21 ›› Issue (1): 357-364.doi: 10.4103/NRR.NRR-D-24-00034

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

促进NSC-34 细胞分化为稳定而持久的神经元:阿糖胞苷的作用

  

  • 出版日期:2026-01-15 发布日期:2025-04-24

Empowering the NSC-34 cell line as a motor neuron model: Cytosine arabinoside’s action

Giuseppe Vitale1, †, Susanna Amadio1 , Francesco Liguori1, 2, Cinzia Volonté1, 2, *   

  1. 1 Experimental Neuroscience and Neurological Disease Models, IRCCS Fondazione Santa Lucia, Rome, Italy;  2 Institute for System Analysis and Computer Science “Antonio Ruberti” (IASI), National Research Council (CNR), Rome, Italy  †Current address: Università Cattolica del Sacro Cuore and IRCCS Fondazione Policlinico Universitario Agostino Gemelli, Largo A. Gemelli 8, 00168 Rome, Italy.
  • Online:2026-01-15 Published:2025-04-24
  • Contact: Cinzia Volonté, PhD, cinzia.volonte@cnr.it.
  • Supported by:
    The present work was supported by FATALSDrug Project [Progetti di Ricerca@CNR SAC.AD002.173.058] from National Research Council, Italy (to CV).

摘要:

NSC-34 细胞系是公认的运动神经元模型。在以往报道的实验条件下,只有NSC-34 细胞系类似于分化后的神经元,但没有表现出广泛和长时间的神经元发生,并在培养过程中保持复制能力。实验旨在NSC-34 细胞中培养一种后有丝分裂表型,以优化持续稳定的神经元发生。结果发现,抗有丝分裂药物阿糖胞苷能促进NSC-34 细胞分化为稳健而持久的神经元,该神经元具有丰富生长锥的长而相互连接的神经元突起,并在培养过程中持续至少6周。此外,阿糖胞苷促进了NSC-34 细胞的神经元启动和移位后的再生,而且大多是不可逆的。阿糖胞苷抑制了细胞增殖抗原 Ki67 的表达,而增强了神经元标志物生长相关蛋白43和神经前体细胞波形蛋白和运动神经元特异性标志物 p75,Islet2,Hb9的表达。总之,这些证据证明了 NSC-34 细胞作为运动神经元模型的合理性,且阿糖胞苷增强了该合理性,从而使该模型能更好地应用于神经退行性病变机制研究。

https://orcid.org/0000-0001-7362-8307 (Cinzia Volonté)

关键词: 阿糖胞苷, NSC-34, 运动神经元, 有丝分裂抑制, 神经元启动, 神经元再生, 神经元分化, Ki67

Abstract: The NSC-34 cell line is a widely recognized motor neuron model and various neuronal differentiation protocols have been exploited. Under previously reported experimental conditions, only part of the cells resemble differentiated neurons; however, they do not exhibit extensive and time-prolonged neuritogenesis, and maintain their duplication capacity in culture. The aim of the present work was to facilitate long-term and more homogeneous neuronal differentiation in motor neuron–like NSC-34 cells. We found that the antimitotic drug cytosine arabinoside promoted robust and persistent neuronal differentiation in the entire cell population. Long and interconnecting neuronal processes with abundant growth cones were homogeneously induced and were durable for up to at least 6 weeks in culture. Moreover, cytosine arabinoside was permissive, dispensable, and mostly irreversible in priming NSC-34 cells for neurite initiation and regeneration after mechanical dislodgement. Finally, the expression of the cell proliferation antigen Ki67 was inhibited by cytosine arabinoside, whereas the expression levels of neuronal growth associated protein 43, vimentin, and motor neuron–specific p75, Islet2, homeobox 9 markers were upregulated, as confirmed by western blot and/or confocal immunofluorescence analysis. Overall, these findings support the use of NSC-34 cells as a motor neuron model for properly investigating neurodegenerative mechanisms and prospectively identifying neuroprotective strategies.

Key words: cytosine arabinoside, Islet2, Hb9, Ki67, mitosis inhibition, neurite initiation, neurite regeneration, neuronal differentiation protocol, NSC-34, p75