Evaluation of fibroblast growth factor‑2 on the proliferation of osteogenic potential and protein expression of stem cell spheroids composed of stem cells derived from bone marrow

Tae,Jae‑Yong; Ko,Youngkyung; Park,Jun‑Beom

doi:10.3892/etm.2019.7543

Evaluation of fibroblast growth factor‑2 on the proliferation of osteogenic potential and protein expression of stem cell spheroids composed of stem cells derived from bone marrow

Authors:
- Jae‑Yong Tae
- Youngkyung Ko
- Jun‑Beom Park
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Affiliations: Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea
Published online on: May 3, 2019 https://doi.org/10.3892/etm.2019.7543
Pages: 326-331
Copyright: © Tae et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Fibroblast growth factor‑2 (FGF‑2) is reported to have various functions and is considered a key human mesenchymal stem cell mitogen, often supplemented to increase human mesenchymal stem cell growth rates. The purpose of this study was to evaluate the effects of FGF‑2 on cellular viability and osteogenic differentiation using three‑dimensional cell spheroids of stem cells. Three‑dimensional cell spheroids were fabricated using concave silicon elastomer‑based microwells in the presence of FGF‑2 at concentrations of 0, 30, 60 and 90 ng/ml. Qualitative cellular viability was determined with a confocal microscope, and quantitative cellular viability was evaluated using a Cell Counting Kit‑8 assay. Alkaline phosphatase activity and Alizarin Red S staining were used to assess osteogenic differentiation. Spheroids were well formed in silicon elastomer‑based concave microwells on Day 1. The average spheroid diameters at Day 1 for FGF‑2 at 0, 30, 60 and 90 ng/ml were 202.2±3.0, 206.6±22.6, 208.8±6.8 and 196.6±26.7 µm, respectively (P>0.05). The majority of the cells in the cell spheroids emitted green fluorescence. The relative Cell Counting Kit‑8 assay values for FGF‑2 at 0, 30, 60 and 90 ng/ml at Day 1 were 100.0±5.5, 101.8±8.8, 99.2±4.8 and 103.4±9.6% (P>0.05). The addition of FGF‑2 at 60 ng/ml concentration produced the highest value for alkaline phosphatase activity. Mineralized extracellular deposits were evenly observed in each group, and the highest value was identified for FGF‑2 groups at 60 ng/ml concentration for Alizarin Red S staining. Based on these findings, it was concluded that FGF‑2 may increase alkaline phosphatase activity or Alizarin Red S staining, and further studies are needed to fully elucidate the mechanisms of FGF‑2.

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