Cerebrospinal fluid-contacting neurons (CSF-cNs) act crucial role in chemosensory and mechanosensory function in spinal cord. Recently, CSF-cNs were found to be an immature neuron and may be involved in spinal cord injury recovery. But how to culture it and explore its function in vitro are not reported in previous research. Here, we first reported culture and identification of CSF-cNs in vitro. We first established a protocol for in vitro culture of CSF-cNs from the cervical spinal cord of mice within 24 h after birth. Polycystic kidney disease 2-like 1 (PKD2L1)⁺ cells were isolated by fluorescence-activated cell sorting and expressed the neuron marker β-tubulin III and CSF-cNs marker GABA. Intriguingly, PKD2L1⁺ cells formed neurosphere and expressed neural stem cell markers Nestin, Sox2 and GFAP. Thus, our research provided culture and isolation of CSF-cNs and this facilitate the investigation the CSF-cNs function in vitro.

脑脊液接触神经元（CSF-cN）在脊髓的化学感觉和机械感觉功能中发挥着至关重要的作用。最近，CSF-cNs被发现是一种不成熟的神经元，可能参与脊髓损伤的恢复。但如何在体外培养它并探讨其功能，以往的研究尚未见报道。在这里，我们首次报道了 CSF-cN 的体外培养和鉴定。我们首先建立了出生后 24 小时内从小鼠颈髓中提取 CSF-cN 的体外培养方案。通过荧光激活细胞分选分离多囊肾病 2 样 1 (PKD2L1) ^{+细胞，并表达神经元标记物 β-微管蛋白 III 和 CSF-cNs 标记物 GABA。}有趣的是，PKD2L1 ⁺细胞形成神经球并表达神经干细胞标记物 Nestin、Sox2 和 GFAP。因此，我们的研究提供了 CSF-cNs 的培养和分离，这有助于研究 CSF-cNs 的体外功能。