Basabrain Mohammed S, Zhong Jialin, Luo Haiyun, Liu Junqing, Yi Baicheng, Zaeneldin Ahmed, Koh Junhao, Zou Ting, Zhang Chengfei
Restorative Dental Sciences, Endodontics, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China.
Stomatological Hospital, Southern Medical University, 366 Jiangnan Avenue South, Guangzhou 510280, China.
Bioengineering (Basel). 2022 Oct 22;9(11):604. doi: 10.3390/bioengineering9110604.
Cell-based neural regeneration is challenging due to the difficulty in obtaining sufficient neural stem cells with clinical applicability. Stem cells from apical papilla (SCAPs) originating from embryonic neural crests with high neurogenic potential could be a promising cell source for neural regeneration. This study aimed to investigate whether the formation of 3D spheres can promote SCAPs' neurogenic potential.
Three-dimensional SCAP spheres were first generated in a 256-well agarose microtissue mold. The spheres and single cells were individually cultured on collagen I-coated μ-slides. Cell morphological changes, neural marker expression, and neurite outgrowth were evaluated by confocal microscope, ELISA, and RT-qPCR.
Pronounced morphological changes were noticed in a time-dependent manner. The migrating cells' morphology changed from fibroblast-like cells to neuron-like cells. Compared to the 2D culture, neurite length, number, and the expression of multiple progenitors, immature and mature neural markers were significantly higher in the 3D spheres. BDNF and NGF-β may play a significant role in the neural differentiation of SCAP spheres.
The formation of 3D spheres enhanced the neurogenic potential of SCAPs, suggesting the advantage of using the 3D spheres of SCAPs for treating neural diseases.
由于难以获得具有临床适用性的足够神经干细胞，基于细胞的神经再生具有挑战性。源自胚胎神经嵴且具有高神经发生潜力的根尖乳头干细胞（SCAPs）可能是神经再生的一种有前途的细胞来源。本研究旨在探讨三维球体的形成是否能促进SCAPs的神经发生潜力。
首先在256孔琼脂糖微组织模具中生成三维SCAP球体。将球体和单细胞分别培养在胶原I包被的μ载玻片上。通过共聚焦显微镜、酶联免疫吸附测定（ELISA）和逆转录定量聚合酶链反应（RT-qPCR）评估细胞形态变化、神经标志物表达和神经突生长。
观察到明显的时间依赖性形态变化。迁移细胞的形态从成纤维细胞样细胞转变为神经元样细胞。与二维培养相比，三维球体中的神经突长度、数量以及多种祖细胞、未成熟和成熟神经标志物的表达均显著更高。脑源性神经营养因子（BDNF）和神经生长因子-β（NGF-β）可能在SCAP球体的神经分化中起重要作用。
三维球体的形成增强了SCAPs的神经发生潜力，表明使用SCAPs的三维球体治疗神经疾病的优势。
