Lee Sung-Il, Ko Youngkyung, Park Jun-Beom
Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea.
Exp Ther Med. 2017 Sep;14(3):2434-2438. doi: 10.3892/etm.2017.4813. Epub 2017 Jul 19.
Three-dimensional cell culture systems provide a convenient model for the study of complex cell-cell and cell-matrix interactions in the absence of exogenous substrates. The current study aimed to evaluate the osteogenic differentiation potential of gingiva-derived stem cells cultured in two-dimensional or three-dimensional systems. To the best of our knowledge, the present study is the first to compare the growth of gingiva-derived stem cells in monolayer culture to a three-dimensional culture system with microwells. For three-dimensional culture, gingiva-derived stem cells were isolated and seeded into polydimethylsiloxane-based concave micromolds. Alkaline phosphatase activity and alizarin red S staining assays were then performed to evaluate osteogenesis and the degree of mineralization, respectively. Stem cell spheroids had a significantly increased level of alkaline phosphatase activity and mineralization compared with cells from the two-dimensional culture. In addition, an increase in mineralized deposits was observed with an increase in the loading cell number. The results of present study indicate that gingiva-derived stem cell spheroids exhibit an increased osteogenic potential compared with stem cells from two-dimensional culture. This highlights the potential of three-dimensional culture systems using gingiva-derived stem cells for regenerative medicine applications requiring stem cells with osteogenic potential.
三维细胞培养系统提供了一个方便的模型，用于在无外源底物的情况下研究复杂的细胞-细胞和细胞-基质相互作用。当前的研究旨在评估在二维或三维系统中培养的牙龈来源干细胞的成骨分化潜能。据我们所知，本研究首次比较了牙龈来源干细胞在单层培养与带有微孔的三维培养系统中的生长情况。对于三维培养，分离牙龈来源干细胞并接种到基于聚二甲基硅氧烷的凹形微模具中。然后分别进行碱性磷酸酶活性和茜素红S染色测定，以评估成骨作用和矿化程度。与二维培养的细胞相比，干细胞球的碱性磷酸酶活性水平和矿化程度显著增加。此外，随着接种细胞数量的增加，观察到矿化沉积物增加。本研究结果表明，与二维培养的干细胞相比，牙龈来源的干细胞球表现出更高的成骨潜能。这突出了使用牙龈来源干细胞的三维培养系统在需要具有成骨潜能干细胞的再生医学应用中的潜力。
