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听力受损豚鼠行蜗窗造口术后小鼠内耳祖细胞/干细胞在柯蒂氏器中的移植与存活:初步结果

Transplantation and survival of mouse inner ear progenitor/stem cells in the organ of Corti after cochleostomy of hearing-impaired guinea pigs: preliminary results.

作者信息

Barboza L C M, Lezirovitz K, Zanatta D B, Strauss B E, Mingroni-Netto R C, Oiticica J, Haddad L A, Bento R F

机构信息

Departamento de Otorrinolaringologia (LIM32), Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.

Setor de Vetores Virais, Laboratório de Genética e Cardiologia Molecular, Instituto do Coração, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.

出版信息

Braz J Med Biol Res. 2016;49(4):e5064. doi: 10.1590/1414-431X20155064. Epub 2016 Mar 18.

Abstract

In mammals, damage to sensory receptor cells (hair cells) of the inner ear results in permanent sensorineural hearing loss. Here, we investigated whether postnatal mouse inner ear progenitor/stem cells (mIESCs) are viable after transplantation into the basal turns of neomycin-injured guinea pig cochleas. We also examined the effects of mIESC transplantation on auditory functions. Eight adult female Cavia porcellus guinea pigs (250-350 g) were deafened by intratympanic neomycin delivery. After 7 days, the animals were randomly divided in two groups. The study group (n=4) received transplantation of LacZ-positive mIESCs in culture medium into the scala tympani. The control group (n=4) received culture medium only. At 2 weeks after transplantation, functional analyses were performed by auditory brainstem response measurement, and the animals were sacrificed. The presence of mIESCs was evaluated by immunohistochemistry of sections of the cochlea from the study group. Non-parametric tests were used for statistical analysis of the data. Intratympanic neomycin delivery damaged hair cells and increased auditory thresholds prior to cell transplantation. There were no significant differences between auditory brainstem thresholds before and after transplantation in individual guinea pigs. Some mIESCs were observed in all scalae of the basal turns of the injured cochleas, and a proportion of these cells expressed the hair cell marker myosin VIIa. Some transplanted mIESCs engrafted in the cochlear basilar membrane. Our study demonstrates that transplanted cells survived and engrafted in the organ of Corti after cochleostomy.

摘要

在哺乳动物中,内耳感觉受体细胞(毛细胞)受损会导致永久性感音神经性听力损失。在此,我们研究了出生后小鼠内耳祖细胞/干细胞(mIESCs)移植到新霉素损伤的豚鼠耳蜗基底转后是否能够存活。我们还检测了mIESC移植对听觉功能的影响。8只成年雌性豚鼠(体重250 - 350克)通过鼓室内注射新霉素致聋。7天后,将动物随机分为两组。研究组(n = 4)在培养基中接受LacZ阳性mIESCs移植到鼓阶。对照组(n = 4)仅接受培养基。移植后2周,通过听觉脑干反应测量进行功能分析,然后处死动物。通过对研究组耳蜗切片进行免疫组织化学评估mIESCs的存在情况。数据采用非参数检验进行统计分析。在细胞移植前,鼓室内注射新霉素损伤了毛细胞并提高了听觉阈值。个体豚鼠移植前后的听觉脑干阈值无显著差异。在受损耳蜗基底转的所有阶中均观察到一些mIESCs,其中一部分细胞表达毛细胞标记物肌球蛋白VIIa。一些移植的mIESCs植入了耳蜗基底膜。我们的研究表明,在进行耳蜗造口术后,移植的细胞在柯蒂氏器中存活并植入。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6180/4819408/09ac135afaa5/1414-431X-bjmbr-1414-431X20155064-gf001.jpg

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