Matsuoka Akihiro J, Sayed Zafar A, Stephanopoulos Nicholas, Berns Eric J, Wadhwani Anil R, Morrissey Zachery D, Chadly Duncan M, Kobayashi Shun, Edelbrock Alexandra N, Mashimo Tomoji, Miller Charles A, McGuire Tammy L, Stupp Samuel I, Kessler John A
Department of Otolaryngology and Head and Neck Surgery, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, United States of America.
Department of Communication Sciences and Disorders, Northwestern University, Evanston, Illinois, United States of America.
PLoS One. 2017 Dec 28;12(12):e0190150. doi: 10.1371/journal.pone.0190150. eCollection 2017.
The use of human embryonic stem cells (hESCs) for regeneration of the spiral ganglion will require techniques for promoting otic neuronal progenitor (ONP) differentiation, anchoring of cells to anatomically appropriate and specific niches, and long-term cell survival after transplantation. In this study, we used self-assembling peptide amphiphile (PA) molecules that display an IKVAV epitope (IKVAV-PA) to create a niche for hESC-derived ONPs that supported neuronal differentiation and survival both in vitro and in vivo after transplantation into rodent inner ears. A feature of the IKVAV-PA gel is its ability to form organized nanofibers that promote directed neurite growth. Culture of hESC-derived ONPs in IKVAV-PA gels did not alter cell proliferation or viability. However, the presence of IKVAV-PA gels increased the number of cells expressing the neuronal marker beta-III tubulin and improved neurite extension. The self-assembly properties of the IKVAV-PA gel allowed it to be injected as a liquid into the inner ear to create a biophysical niche for transplanted cells after gelation in vivo. Injection of ONPs combined with IKVAV-PA into the modiolus of X-SCID rats increased survival and localization of the cells around the injection site compared to controls. Human cadaveric temporal bone studies demonstrated the technical feasibility of a transmastoid surgical approach for clinical intracochlear injection of the IKVAV-PA/ONP combination. Combining stem cell transplantation with injection of self-assembling PA gels to create a supportive niche may improve clinical approaches to spiral ganglion regeneration.
利用人类胚胎干细胞(hESCs)实现螺旋神经节再生,将需要促进耳神经元祖细胞(ONP)分化、使细胞锚定到解剖学上合适且特定的微环境,以及移植后细胞长期存活的技术。在本研究中,我们使用展示IKVAV表位的自组装肽两亲分子(PA)(IKVAV-PA)为hESC来源的ONP创建一个微环境,该微环境在体外以及移植到啮齿动物内耳后在体内均支持神经元分化和存活。IKVAV-PA凝胶的一个特点是其能够形成有组织的纳米纤维,促进神经突定向生长。在IKVAV-PA凝胶中培养hESC来源的ONP不会改变细胞增殖或活力。然而,IKVAV-PA凝胶的存在增加了表达神经元标志物β-III微管蛋白的细胞数量,并改善了神经突延伸。IKVAV-PA凝胶的自组装特性使其能够作为液体注入内耳,以便在体内凝胶化后为移植细胞创建一个生物物理微环境。与对照组相比,将ONP与IKVAV-PA联合注入X-SCID大鼠的蜗轴可提高细胞在注射部位周围的存活率和定位。人类尸体颞骨研究证明了经乳突手术方法用于临床耳蜗内注射IKVAV-PA/ONP组合的技术可行性。将干细胞移植与注射自组装PA凝胶以创建支持性微环境相结合,可能会改善螺旋神经节再生的临床方法。