毒性损伤大鼠耳蜗核移植嗅球神经前体细胞的初步观察

目的体外培养大鼠嗅球神经前体细胞，并移植到谷氨酸诱导损伤的耳蜗核，观察其生存、分化过程。方法嗅球神经前体细胞取自孕15d胚胎大鼠，免疫荧光染色鉴定。耳蜗核定位注射谷氨酸制成损伤模型。伤后7d移植标记的神经前体细胞，不同时间测定听性脑干反应并取材观察移植细胞的存活及分化。结果巢蛋白阳性的嗅球神经前体细胞在体外可自我复制传代，并分化出神经元和神经胶质细胞。谷氨酸损伤和前体细胞移植均造成听性脑干诱发反应（ABR）阈值升高，并有部分的恢复。免疫荧光技术发现耳蜗核中Hoechst33342分别和神经元特异性烯醇化酶（NSE）、胶质纤维酸性蛋白（GFAP）、谷氨酸双重标记的移植细胞。结论嗅球神经前体细胞移植耳蜗核短期存活良好，所分化出的神经元可表达听觉神经递质。

Olfactory bulb derived neural precursor cells transplantation in toxic injured cochlear nucleus of rats

Objective In vitro culturing neural precursor cells (NPCs) from embryonic rat olfactory bulb and observing the survival and differentiation of the NPCs transplanted into injured cochlear nucleus of rats induced by injecting glutamate. Methods NPCs from 15 days embryonic rat olfactory bulb were cultured and identified with immunofluorescence technique. Stereotaxic injection of glutamic acid established a cochlear nucleus impairment model. The NPCs labeled with hoechst33342 were transplanted 7 days after toxicity impairment. The survival and differentiation of labeled cells were observed up to 6 weeks. Acoustic brainstem evoked response (ABR) were tested at different time.Results Nestin-positive NPCs with self-renewal capacity could differentiate into neurons and astrocytes. Both glutamate toxicity and the injection of NPCs elevated ABR thresholds, which got back partially later. Double-labeled cells by Hoechst33342 respectively with neuron specific enolase (NSE), glial fibrillary acid protein (GFAP), as well as glutamate were found in the cochlear nuclei with immunofluorescence technique. Conclusion NPCs from olfactory bulb can survive in cochlear nucleus and differentiate into neurons expressing auditory neurotransmitter.