中国神经再生研究(英文版) ›› 2016, Vol. 11 ›› Issue (6): 994-1000.doi: 10.4103/1673-5374.184503

• 原著:颅神经损伤修复保护与再生 • 上一篇    下一篇

人脂肪干细胞诱导分化的神经元样细胞移植修复耳聋豚鼠螺旋神经节神经元

  

  • 收稿日期:2015-12-22 出版日期:2016-06-30 发布日期:2016-06-30
  • 基金资助:

    韩国教育科技部基础研究项目(2010-0025501);Chonnam国立大学医院科研项目(2012-2894)

Transplantation of human adipose tissue-derived stem cells for repair of injured spiral ganglion neurons in deaf guinea pigs

Sujeong Jang1, *, Hyong-Ho Cho2, Song-Hee Kim1, Kyung-Hwa Lee3, Yong-Bum Cho2, Jong-Seong Park1, Han-Seong Jeong1,*   

  1. 1 Department of Physiology, Chonnam National University Medical School, Gwangju, Republic of Korea 2 Department of Otolaryngology, Chonnam National University Medical School, Gwangju, Republic of Korea 3 Department of Pathology, Chonnam National University Medical School, Gwangju, Republic of Korea
  • Received:2015-12-22 Online:2016-06-30 Published:2016-06-30
  • Contact: Han-Seong Jeong, M.D., Ph.D. or Sujeong Jang, Ph.D., jhsjeong@hanmail.net or sujeong.jjang@gmail.com.
  • Supported by:

    This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by Ministry of Education, Science and Technology, No. 2010-0025501; a grant from Chonnam National University Hospital Biomedical Research Institute, No. CRI11048-1 and a grant from the Chonnam National University, No. 2012-2894.

摘要:

噪音、耳毒性药物、感染等原因可导致螺旋神经节神经元丢失,使哺乳动物发生感音性耳聋。研究已证实干细胞可分化为螺旋神经节神经元,但目前尚缺乏脂肪干细胞用于修复损伤螺旋神经节神经元的研究。我们假设人脂肪干细胞诱导分化的神经元样细胞移植至豚鼠新霉素损害的内耳可修复损伤的螺旋神经节神经元。实验应用添加碱性成纤维生长因子和forskolin神经诱导培养基诱导人脂肪干细胞向神经元样细胞分化,然后将其注射至损伤的耳蜗;对照组为以损伤耳蜗内注射Hank's平衡盐溶液的豚鼠。细胞移植后8周,与对照组比较,HE染色显示存活螺旋神经节神经元数量明显增加;免疫组化染色显示螺旋神经节区检测到更多的移植的人脂肪干细胞诱导分化的神经元样细胞,这些细胞表达神经元标志物神经丝蛋白和微管相关蛋白2。细胞移植后8周内,80dB短声和纯音刺激诱发的听性脑干反应阈值呈下降趋势提示,而对照组豚鼠则对80dB的短声和纯音刺激无反应。说明大量人脂肪干细胞诱导分化的神经元样细胞迁移至螺旋神经节,并较长时间存活,修复其损伤,从而促进感音性耳聋豚鼠听力的恢复。 

orcid: 0000-0001-6921-6625 (Han-Seong Jeong) 0000-0001-8673-7887 (Sujeong Jang)

关键词: 神经再生, 脂肪干细胞, 螺旋神经节, 神经元, 神经分化, 干细胞移植, 听力损失, 脑干听觉诱发电位

Abstract:

Excessive noise, ototoxic drugs, infections, autoimmune diseases, and aging can cause loss of spiral ganglion neurons, leading to permanent sensorineural hearing loss in mammals. Stem cells have been confirmed to be able to differentiate into spiral ganglion neurons. Little has been reported on adipose tissue-derived stem cells (ADSCs) for repair of injured spiral ganglion neurons. In this study, we hypothesized that transplantation of neural induced-human ADSCs (NI-hADSCs) can repair the injured spiral ganglion neurons in guinea pigs with neomycin-induced sensorineural hearing loss. NI-hADSCs were induced with culture medium containing basic fibroblast growth factor and forskolin and then injected to the injured cochleae. Guinea pigs that received injection of Hanks’ balanced salt solution into the cochleae were used as controls. Hematoxylin-eosin staining showed that at 8 weeks after cell transplantation, the number of surviving spiral ganglion neurons in the cell transplantation group was significantly increased than that in the control group. Also at 8 weeks after cell transplantation, immunohistochemical staining showed that a greater number of NI-hADSCs in the spiral ganglions were detected in the cell transplantation group than in the control group, and these NI-hADSCs expressed neuronal markers neurofilament protein and microtubule-associated protein 2. Within 8 weeks after cell transplantation, the guinea pigs in the cell transplantation group had a gradually decreased auditory brainstem response threshold, while those in the control group had almost no response to 80 dB of clicks or pure tone burst. These findings suggest that a large amount of NI-hADSCs migrated to the spiral ganglions, survived for a period of time, repaired the injured spiral ganglion cells, and thereby contributed to the recovery of sensorineural hearing loss in guinea pigs.

Key words: nerve regeneration, adipose tissue-derived stem cells, spiral ganglion, neurons, hearing impairment, stem cell transplantation, brainstem auditory evoked potential, neural differentiation