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Evaluation of the maintenance of stemness, viability, and differentiation potential of gingiva-derived stem-cell spheroids

  • Authors:
    • Sung‑Il Lee
    • Youngkyung Ko
    • Jun‑Beom Park
  • View Affiliations / Copyright

    Affiliations: Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea
    Copyright: © Lee et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 1757-1764
    |
    Published online on: March 8, 2017
       https://doi.org/10.3892/etm.2017.4194
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Abstract

Gingiva-derived stem cells have been applied for tissue‑engineering purposes and may be considered a favorable source of mesenchymal stem cells as harvesting stem cells from the mandible or maxilla may be performed with ease under local anesthesia. The present study was performed to fabricate stem‑cell spheroids using concave microwells and to evaluate the maintenance of stemness, viability, and differentiation potential. Gingiva‑derived stem cells were isolated, and the stem cells of 4x105 (group A) or 8x105 (group B) cells were seeded into polydimethylsiloxane‑based, concave micromolds with 600 µm diameters. The morphology of the microspheres and the change of the diameters of the spheroids were evaluated. The viability of spheroids was qualitatively analyzed via Live/Dead kit assay. A cell viability analysis was performed on days 1, 3, 6, and 12 with Cell Counting Kit‑8. The maintenance of stemness was evaluated with immunocytochemical staining using SSEA‑4, TRA‑1‑60(R) (positive markers), and SSEA‑1 (negative marker). Osteogenic, adipogenic, and chondrogenic differentiation potential was evaluated by incubating spheroids in osteogenic, adipogenic and chondrogenic induction medium, respectively. The gingiva‑derived stem cells formed spheroids in the concave microwells. The diameters of the spheroids were larger in group A than in group B. The majority of cells in the spheroids emitted green fluorescence, indicating the presence of live cells at day 6. At day 12, the majority of cells in the spheroids emitted green fluorescence, and a small portion of red fluorescence was also noted, which indicated the presence of dead cells. The spheroids were positive for the stem‑cell markers SSEA‑4 and TRA‑1‑60(R) and were negative for SSEA‑1, suggesting that these spheroids primarily contained undifferentiated human stem cells. Osteogenic, adipogenic, and chondrogenic differentiation was more evident with an increase of incubation time: Mineralized extracellular deposits were observed following Alizarin Red S staining at days 14 and 21; oil globules were increased at day 18 when compared with day 6; and Alcian blue staining was more evident at day 18 when compared with day 6. Within the limits of this study, stem‑cell spheroids from gingival cells maintained the stemness, viability, and differentiation potential during the experimental periods. This method may be applied for a promising strategy for stem-cell therapy.
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Copy and paste a formatted citation
Spandidos Publications style
Lee SI, Ko Y and Park JB: Evaluation of the maintenance of stemness, viability, and differentiation potential of gingiva-derived stem-cell spheroids. Exp Ther Med 13: 1757-1764, 2017.
APA
Lee, S., Ko, Y., & Park, J. (2017). Evaluation of the maintenance of stemness, viability, and differentiation potential of gingiva-derived stem-cell spheroids. Experimental and Therapeutic Medicine, 13, 1757-1764. https://doi.org/10.3892/etm.2017.4194
MLA
Lee, S., Ko, Y., Park, J."Evaluation of the maintenance of stemness, viability, and differentiation potential of gingiva-derived stem-cell spheroids". Experimental and Therapeutic Medicine 13.5 (2017): 1757-1764.
Chicago
Lee, S., Ko, Y., Park, J."Evaluation of the maintenance of stemness, viability, and differentiation potential of gingiva-derived stem-cell spheroids". Experimental and Therapeutic Medicine 13, no. 5 (2017): 1757-1764. https://doi.org/10.3892/etm.2017.4194
Copy and paste a formatted citation
x
Spandidos Publications style
Lee SI, Ko Y and Park JB: Evaluation of the maintenance of stemness, viability, and differentiation potential of gingiva-derived stem-cell spheroids. Exp Ther Med 13: 1757-1764, 2017.
APA
Lee, S., Ko, Y., & Park, J. (2017). Evaluation of the maintenance of stemness, viability, and differentiation potential of gingiva-derived stem-cell spheroids. Experimental and Therapeutic Medicine, 13, 1757-1764. https://doi.org/10.3892/etm.2017.4194
MLA
Lee, S., Ko, Y., Park, J."Evaluation of the maintenance of stemness, viability, and differentiation potential of gingiva-derived stem-cell spheroids". Experimental and Therapeutic Medicine 13.5 (2017): 1757-1764.
Chicago
Lee, S., Ko, Y., Park, J."Evaluation of the maintenance of stemness, viability, and differentiation potential of gingiva-derived stem-cell spheroids". Experimental and Therapeutic Medicine 13, no. 5 (2017): 1757-1764. https://doi.org/10.3892/etm.2017.4194
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