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目的观察未经体外诱导的胚胎干细胞(embyonic stem cells,ESC)导入听力正常大鼠内耳的可行性以及导入后的存活和分布情况,为ESC内耳移植治疗由毛细胞缺失导致的感音神经性耳聋提供实验基础和理论依据。方法5～6周龄Wistar大鼠,10只,右耳为实验耳:经鼓阶打孔法植入带有绿色荧光蛋白(EGFP)的ESCs;左耳为对照组,不实施手术。术前1周与术后即刻行听性脑干反应(ABR)检查,取双侧耳蜗做冰冻切片,观察ESCs植入内耳后存活和分布情况。结果术后动物存活8只,麻醉效果好无干扰完整测完ABR动物5只。鼓阶打孔途径导入耳蜗的ESCs大部分于鼓阶聚集悬浮,少数可在鼓阶基底膜嵴和鼓阶外侧壁处贴壁;未在柯替器等中阶部位中发现有ESCs的分布。ABR检测结果显示鼓阶打孔途径导入方法对大鼠听力影响较小。结论胚胎干细胞可经耳蜗底转鼓阶打孔途径导入耳蜗,干细胞在内耳成功存活,并且对内耳损伤小,因此,它可以作为内耳细胞移植的重要方式。
Objective To observe the feasibility of introduction of embryonic stem cells (ESC) into the inner ear of normal rats without inducing in vitro and the survival and distribution after implantation. To evaluate the effect of ESC inner ear transplantation on sensorineural hearing loss caused by hair loss Provide experimental basis and theoretical basis. Methods Wistar rats (5 ~ 6 weeks old), 10 ears were used as the experimental ear. ESCs with green fluorescence protein (EGFP) were implanted by the scala tympani implantation. The left ear was the control group. One week before and immediately after operation, auditory brainstem response (ABR) was performed. The bilateral cochleas were frozen sectioned to observe the survival and distribution of ESCs after implantation into the inner ear. Results Postoperative animals survived 8, with good anesthetic effect without interfering with the complete measurement of ABR animals 5. Most of the ESCs introduced into the cochlea of the cochlear implants were aggregated and suspended in the scala tympani. A few of them adhered to the basal lamina of the basilar membrane and the outer wall of the scala tympani. ESCs were not found in the midcourse of the cochlear implant. ABR test results showed that the method of inducting drum-hole perforation had little effect on hearing in rats. Conclusion Embryonic stem cells can be introduced into the cochlea via the cochlear inflow drum. The stem cells survive successfully in the inner ear and have less damage to the inner ear. Therefore, it can be used as an important way for transplanting the inner ear cells.