Abstract: Objective　To explore the isolation, purification and in vitro culture conditions of naked mole rat neural stem cells and the effects of 60Co-γ rays on naked mole rat neural stem cells.Methods　The subventricular zone (SVZ) brain tissue of fetuses from 8-9-week pregnant naked mole rat was isolated, and an in vitro culture system of naked mole rat neural stem cells was established. The isolated naked mole rat neural stem cells were purified based on their physical characteristics, specific SRY-box transcription factor 2(SOX2)immunofluorescence labeling and multi-directional differentiation potential. Subsequently, the in vitro cultured naked mole rat neural stem cells were irradiated with 10 Gy 60Co-γ rays in a single dose. The comet assay, flow cytometry cell cycle assay, qPCR assay and neurosphere assay were used to investigate the effects of irradiation on DNA damage, proliferation and growth of naked mole rat neural stem cells.Results　The cultured naked mole rat neural stem cells had the characteristic of spherical growth of stem cells, could specifically express neural stem cell-specific markers, and had good multi-directional differentiation potential. The growth cycle of naked mole rat neural stem cells was slow. After irradiation, they could repair the DNA damage within a short time and maintain a normal growth rate and state, indicating that naked mole rat neural stem cells have certain radiation stability. Conclusion　The in vitro growth of naked mole rat neural stem cells has special environmental requirements, with a low growth rate, and naked mole rat neural stem cells have strong radiation resistance.

Key words: naked mole-rat, neural stem cells, radiation, proliferation, differentiation, DNA damage

摘要： 目的　探究裸鼹鼠神经干细胞分离纯化和体外培养条件以及60Co-γ射线对裸鼹鼠神经干细胞的影响。方法 取孕8~9周的裸鼹鼠胎鼠脑室下区(SVZ)脑组织,对其进行分离,建立裸鼹鼠神经干细胞体外培养体系。根据干 细胞物理特性、特殊的SRY-box转录因子2(SOX2)免疫荧光标记和多向分化潜能鉴定纯化所得的裸鼹鼠神经干细 胞。随后利用10 Gy 60Co-γ射线对体外培养的裸鼹鼠神经干细胞一次性照射,利用彗星实验、流式细胞周期实验、 实时荧光定量PCR(qPCR)方法实验以及神经球测定探究辐照对裸鼹鼠神经干细胞DNA损伤、增殖和生长的影 响。结果　所培养的裸鼹鼠神经干细胞具有干细胞的成球生长特性,能特异性表达神经干细胞专属标志物,且具 有良好的多向分化潜能。裸鼹鼠神经干细胞生长周期缓慢,辐照后能在短时间内修复受到损伤的DNA,并保持正 常的生长速率和状态,表明裸鼹鼠神经干细胞具有一定的辐射稳定性。结论　裸鼹鼠神经干细胞体外生长具有特 殊的环境要求,生长速率较慢,且裸鼹鼠神经干细胞具有较强的抗辐射特性。

关键词: 裸鼹鼠, 神经干细胞, 辐射, 增殖, 分化, DNA损伤