Wu Yen, Chung Yao-Yu, Chin Yu-Tang, Lin Chi-Yu, Kuo Po-Jan, Chen Ting-Yi, Lin Tzu-Yu, Chiu Hsien-Chung, Huang Haw-Ming, Jeng Jiiang-Huei, Lee Sheng-Yang
School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei, Taiwan.
Department of Dentistry, Wan-Fang Medical Center, Taipei Medical University, Taipei, Taiwan.
J Dent Sci. 2022 Jan;17(1):14-29. doi: 10.1016/j.jds.2021.09.021. Epub 2021 Sep 24.
BACKGROUND/PURPOSE: Culture environments play a critical role in stem cell expansion. This study aimed to evaluate the effects of 2,3,5,4'-tetrahydroxystilbene-2-O-b-D-glucoside (THSG) on the proliferation and differentiation of human dental pulp stem cells (DPSCs) in 2-dimensional (2D) and 3-dimensional (3D) culture systems.
Human DPSCs were seeded in T25 flasks for 2D cultivation. For the 3D culture system, DPSCs were mixed with microcarriers and cultured in spinner flasks. Cells in both culture systems were treated with THSG, and cell proliferation was determined using a cell counter and a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. In THSG-treated DPSCs, the genes associated with proliferation, adipogenesis, neurogenesis, osteogenesis, pluripotency, oncogenesis, and apoptosis were analyzed using real-time polymerase chain reactions.
The spinner flask time-dependently improved cell numbers, cell viability, and expansion rates in THSG-treated DPSCs. In both the T25 and spinner flasks, the messenger RNA (mRNA) levels of proliferation, osteogenesis, and pluripotent-related genes had a significant maximum expression with 10 μM THSG treatment. However, 0.1 μM of THSG may be the most suitable condition for triggering neurogenesis and adipogenesis gene expression when DPSCs were cultured in spinner flasks. Furthermore, the number of oncogenes and apoptotic genes decreased considerably in the presence of THSG in both the T25 and spinner flasks.
The spinner flask bioreactor combined with THSG may upregulate proliferation and lineage-specific differentiation in DPSCs. Thus, the combination can be used to mass-produce and cultivate human DPSCs for regenerative dentistry.
背景/目的：培养环境在干细胞扩增中起着关键作用。本研究旨在评估2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷（THSG）在二维（2D）和三维（3D）培养系统中对人牙髓干细胞（DPSC）增殖和分化的影响。
将人DPSC接种于T25培养瓶中进行二维培养。对于三维培养系统，将DPSC与微载体混合并在转瓶中培养。两个培养系统中的细胞均用THSG处理，使用细胞计数器和3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑法测定细胞增殖。在经THSG处理的DPSC中，使用实时聚合酶链反应分析与增殖、脂肪生成、神经生成、成骨、多能性、肿瘤发生和凋亡相关的基因。
转瓶在时间上依赖性地提高了经THSG处理的DPSC的细胞数量、细胞活力和扩增率。在T25培养瓶和转瓶中，增殖、成骨和多能相关基因的信使核糖核酸（mRNA）水平在10 μM THSG处理时均有显著的最大表达。然而，当DPSC在转瓶中培养时，0.1 μM的THSG可能是触发神经生成和脂肪生成基因表达的最合适条件。此外，在T25培养瓶和转瓶中，THSG存在时癌基因和凋亡基因的数量显著减少。
转瓶生物反应器与THSG联合使用可能上调DPSC的增殖和谱系特异性分化。因此，该组合可用于大规模生产和培养用于再生牙科的人DPSC。
