Wei Na, Liu Ya-Ping, Wang Rui-Rui, Zhong Zhen-Juan, Wang Xiao-Liang, Yang Yan, He Ting, Zhao Si-Jia, Wang Huan, Yu Yao-Qing
Institute for Biomedical Sciences of Pain, Tangdu Hospital, Fourth Military Medical University, 569 Xinsi Road, Baqiao, Xi'an, Shaanxi, 710038, People's Republic of China.
Department of Dermatology, Tangdu Hospital, Fourth Military Medical University, 569 Xinsi Road, Baqiao, Xi'an, 710038, People's Republic of China.
Neurochem Res. 2022 Dec;47(12):3635-3646. doi: 10.1007/s11064-022-03614-z. Epub 2022 May 6.
Satellite glial cells (SGCs) tightly surround neurons and modulate sensory transmission in dorsal root ganglion (DRG). At present, the biological property of primary SGCs in culture deserves further investigation. To reveal the key factor for SGCs growth and survival, we examined the effects of different culture supplementations containing Dulbecco's Modified Eagle Medium (DMEM)/F12, DMEM high glucose (HG) or Neurobasal-A (NB). CCK-8 proliferation assay showed an increased proliferation of SGCs in DMEM/F12 and DMEM/HG, but not in NB medium. Bax, AnnexinV, and propidium iodide (PI) staining results showed that NB medium caused cell death and apoptosis. We showed that glutamine was over 2.5 mM in DMEM/F12 and DMEM/HG, whereas it was absence in NB medium. Interestingly, exogenous glutamine application significantly reversed the poor proliferation and cell death of SGCs in NB medium. These findings demonstrated that DMEM/F12 medium was optimal to get high-purity SGCs. Glutamine was the key molecule to maintain SGCs growth and survival in culture. Here, we provided a novel approach to get high-purity SGCs by changing the key component of culture medium. Our study shed a new light on understanding the biological property and modulation of glial cells of primary sensory ganglia.
卫星神经胶质细胞（SGCs）紧密围绕神经元，并调节背根神经节（DRG）中的感觉传递。目前，培养的原代SGCs的生物学特性值得进一步研究。为了揭示SGCs生长和存活的关键因素，我们研究了含有杜氏改良 Eagle 培养基（DMEM）/F12、高糖 DMEM（HG）或 Neurobasal-A（NB）的不同培养基添加剂的作用。CCK-8增殖试验表明，DMEM/F12和DMEM/HG中SGCs的增殖增加，但在NB培养基中没有增加。Bax、膜联蛋白V和碘化丙啶（PI）染色结果表明，NB培养基导致细胞死亡和凋亡。我们发现，DMEM/F12和DMEM/HG中的谷氨酰胺超过2.5 mM，而NB培养基中没有谷氨酰胺。有趣的是，外源性谷氨酰胺的应用显著逆转了NB培养基中SGCs增殖不良和细胞死亡的情况。这些发现表明，DMEM/F12培养基是获得高纯度SGCs的最佳选择。谷氨酰胺是维持培养中SGCs生长和存活的关键分子。在这里，我们提供了一种通过改变培养基关键成分来获得高纯度SGCs的新方法。我们的研究为理解初级感觉神经节胶质细胞的生物学特性和调节提供了新的思路。
