Comparison of 2- and 3-Dimensional Cultured Periodontal Ligament Stem Cells; a Pilot Study

Abstract

This study compared the characteristics of periodontal ligament stem cells (PDLSCs) cultured using 3-dimensional (3D) versus conventional 2-dimensional (2D) methods. PDLSCs were cultured in either a 3D culture with a non-adhesive culture plate (Stemfit 3D®) or a conventional 2D culture using a 6-well plate. Morphology, viability, proliferation ability, and osteogenic differentiation were analyzed to characterize the differences induced in identical PDLSCs by 3D and 2D culture environments. In addition, gene expression was analyzed using RNA sequencing to further characterize the functional differences. The diameter and the viability of the 3D-cultured PDLSCs decreased over time, but the shape of the spheroid was maintained for 20 days. Although osteogenic differentiation occurred in both the 2D- and 3D-cultured PDLSCs, compared to the control group it was 20.8 and 1.6 higher in the 3D- and 2D-cultured cells, respectively. RNA sequencing revealed that PDLSCs cultured using 2D and 3D methods have different gene expression profiles. The viability of the 3D-cultured cells was decreased, but they showed superior osteogenic differentiation compared to 2D-cultured cells. Within the limitations of this study, the results demonstrate that the structure and function of PDLSCs are influenced by the cell culture method.

Brain Homogenate of a Rat Model of Alzheimer's Disease Modifies the Secretome of 3D Cultured Periodontal Ligament Stem Cells: A Potential Neuroregenerative Therapy

“…We clearly demonstrated that the 3D cultured spheroids of isolated PDLSCs were fully formed after 7 days of culturing in a non-adherent environment, and the viability of the cells in 3D cultured spheroids was confirmed at different time points of the spheroid formation. Our results contrast with the results reported by Jeong et al showing that the viability of PDLSCs in 3D culturing decreased compared to traditional 2D culturing ( 40 ). Besides, based on the culturing method, the spheroid morphology was preserved for 3 - 21 days, and the internal structure was homogeneous and biologically stable for at least 30 days ( 41 , 42 ).…”

Anti-Inflammatory Potential of Complex Extracts of Ligularia stenocephala Matsum. & Koidz. and Secale cereale L. Sprout in Chronic Gingivitis: In Vitro Investigation and Randomized Clinical Trial

“…Human periodontal ligament (PDL) cell (CEFOgro™, Seoul, Korea) was cultured according to the manufacturer's protocol [17]. In brief, the cells were cultured in the growth medium containing 100 U/mL penicillin and 100 µg/mL streptomycin.…”

In Vitro Characterization of Periodontal Ligament Stem Cells Derived from Supernumerary Teeth in Three-Dimensional Culture Method

“…Fourth, several prior studies on spheroid morphology involved observations over only 5-7 days. In the previous work of this research team, pPDLSCs were observed for 20 days to confirm their long-term stability [9]. However, the long-term stability of sPDLSC spheroids was not evaluated in this study.…”

“…Periodontal ligament stem cells (PDLSCs) are easy to obtain and have a greater proliferation rate than bone marrow stem cells. PDLSCs are capable of differentiating into osteoblasts, cementoblasts, adipocytes, and chondrocytes, and can form a structure similar to periodontal ligaments in vitro [6][7][8][9].…”