Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Karolinska University Hospital, Huddinge, Sweden.
Department for Surgical Sciences, ENT-Unit, Akademiska Hospital, Uppsala, Sweden.
Anat Rec (Hoboken). 2020 Mar;303(3):441-450. doi: 10.1002/ar.24131. Epub 2019 May 8.
We have previously developed an in vitro organotypic culture setting in order to investigate the performance of cellular substrates transplanted to the auditory nervous system. We have utilized this system to predict the efficacy of human neural progenitor cells (HNPCs) in transplantation to the auditory nerve to facilitate regeneration of sensory auditory nerve structures in vivo and in vitro. To optimize the growth and differentiation of HNPCs we have introduced an expansion of our in vitro system, exploring the impact of a growth factor-altered microenvironment. Here, we seeded HNPCs as a dissociated sphere culture on a hydrogel matrix coating (Matrigel®). We evaluated the performance of HNPCs by studying their survival, differentiation, and their axon-forming capacity. In identical culture conditions, we found that the overall survival rate of HNPCs on Matrigel coated surfaces was better than that on surfaces that were not coated with Matrigel. Furthermore, cells on Matrigel differentiated into neuronal cells to a far greater extent leading to strong synaptic marker signatures. Overall, our findings show that the present Matrigel matrix setting offers an experimental environment for the HNPCs to grow where these cells show novel and promising phenotypic characteristics suitable for further in vivo transplantation to the auditory nerve. Anat Rec, 303:441-450, 2020. © 2019 American Association for Anatomy.
我们之前开发了一种体外器官型培养体系,以研究移植到听觉神经系统的细胞基质的性能。我们利用该系统来预测人神经祖细胞(HNPCs)在移植到听神经中的疗效,以促进体内和体外感觉听觉神经结构的再生。为了优化 HNPCs 的生长和分化,我们引入了体外系统的扩展,探索了生长因子改变的微环境的影响。在这里,我们将 HNPCs 作为一个分离的球体培养物接种在水凝胶基质涂层(Matrigel®)上。我们通过研究 HNPCs 的存活、分化和轴突形成能力来评估它们的性能。在相同的培养条件下,我们发现 HNPCs 在涂有 Matrigel 的表面上的总体存活率优于未涂有 Matrigel 的表面上的存活率。此外,Matrigel 上的细胞分化为神经元细胞的程度要高得多,导致强烈的突触标记特征。总的来说,我们的研究结果表明,目前的 Matrigel 基质设置为 HNPCs 的生长提供了一个实验环境,这些细胞表现出新颖且有前途的表型特征,适合进一步移植到听神经。解剖学年鉴,303:441-450,2020。©2019 美国解剖学会。