Lee Tzu-Pei, Ho Ming-Hua, Chen Yong-Ren, Lin Kuan-Ming, Li Chung-Hsing
Graduate Institute of Dental Science, National Defense Medical Center, Taipei, Taiwan.
Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan.
J Dent Sci. 2025 Jul;20(3):1629-1638. doi: 10.1016/j.jds.2025.03.008. Epub 2025 Mar 20.
BACKGROUND/PURPOSE: Retinopathy affects millions worldwide with increasing prevalence. Dental-derived mesenchymal stem cells (MSCs), primarily originating from the neural crest, possess unique neural-like properties, including high expression of neural markers, making them promising candidates for neural and retinal regenerative medicine. The present study compared two isolation methods for dental-derived MSCs and investigated the potential of dental follicle stem cells (DFSCs) to differentiate into neurons and retinal progenitor cells under defined conditions.
The cells of extracted dental tissues were isolated by enzymatic digestion and explant culture methods. The multi-lineage differentiation capabilities of DFSCs were confirmed by Alizarin Red staining and Oil Red-O staining after defined culture stimulation. The differentiation of neurons and retinal progenitors was induced in DFSCs and confirmed through cell morphology, immunofluorescence staining, and western blotting.
The enzymatic digestion method was faster and promoted quicker proliferation of DFSCs compared to other dental sources. The expression of Alizarin Red staining and Oil Red-O staining confirmed the multi-lineage differentiation capabilities of DFSCs. Under specific conditions, DFSCs demonstrated the potential to differentiate into neural, retinal progenitor, ganglion, bipolar, amacrine, and Müller-like cells with morphologic changes and expression of related proteins.
The present study demonstrates the superior proliferation ability of DFSCs among dental cells, and possesses the potential to differentiate into retinal progenitor cells and neurons under defined conditions. Provide valuable insights into DFSCs as an effective stem cell source and their potential for future therapeutic applications in retinal-related diseases.
背景/目的：视网膜病变在全球影响着数百万人，且患病率不断上升。源自牙源性的间充质干细胞（MSCs）主要起源于神经嵴，具有独特的神经样特性，包括神经标志物的高表达，使其成为神经和视网膜再生医学中有前景的候选细胞。本研究比较了两种牙源性MSCs的分离方法，并研究了牙囊干细胞（DFSCs）在特定条件下分化为神经元和视网膜祖细胞的潜力。
通过酶消化和组织块培养法分离提取的牙组织细胞。在特定培养刺激后，通过茜素红染色和油红O染色确认DFSCs的多向分化能力。诱导DFSCs向神经元和视网膜祖细胞分化，并通过细胞形态学、免疫荧光染色和蛋白质印迹法进行确认。
与其他牙源性来源相比，酶消化法更快，且促进了DFSCs的更快增殖。茜素红染色和油红O染色的表达证实了DFSCs的多向分化能力。在特定条件下，DFSCs表现出分化为神经细胞、视网膜祖细胞、神经节细胞、双极细胞、无长突细胞和穆勒样细胞的潜力，伴有形态学变化和相关蛋白的表达。
本研究证明了DFSCs在牙源性细胞中具有卓越的增殖能力，并且在特定条件下具有分化为视网膜祖细胞和神经元的潜力。为DFSCs作为一种有效的干细胞来源及其在视网膜相关疾病未来治疗应用中的潜力提供了有价值的见解。
