Tissue Eng Regen Med.  2016 Apr;13(2):162-170. 10.1007/s13770-016-9134-x.

Expression of Tenocyte Lineage-Related Factors from Tonsil-Derived Mesenchymal Stem Cells

Affiliations
  • 1Department of Molecular Medicine, School of Medicine, Ewha Womans University, Seoul, Korea. inhojo@ewha.ac.kr
  • 2Department of Orthopaedic Surgery, School of Medicine, Ewha Womans University, Seoul, Korea. sjshin622@ewha.ac.kr
  • 3Ewha Tonsil-derived Mesenchymal Stem Cells Research Center (ETSRC), School of Medicine, Ewha Womans University, Seoul, Korea.

Abstract

Human palatine tonsil-derived mesenchymal stem cells (TMSCs) are known to be a new source of progenitor cells. Using waste tissue after tonsillectomy as a cell provider can be the biggest benefit of TMSCs, compared with other stem cells. The purpose of this study was to investigate tenogenic differentiation of TMSCs and to access the differential effects of transforming growth factor beta 3 (TGF-β3) on the tenogenesis of TMSCs. Human tonsil was obtained after tonsillectomy. Using a cytometric analysis, we were able to find that the TMSCs had typical mesenchymal stem cell markers: positive for CD73, CD90, and CD105, and negative for CD14, CD34, and CD45. Using TGF-β3, the expressions of tenocyte-specific genes and proteins, such as collagen type 1 (COL1), tenomodulin (TNMD), and scleraxis (SCX), were measured by a quantitative polymerase chain reaction (PCR), immunofluorescence staining, immunohistochemistry and Western blot analyses. Quantitative PCR assay showed that TGF-β3 significantly increased the expressions of tenocyte lineage marker genes, including COL1, TNMD, and SCX, at a 3-day treatment, compared with control. However, these increases were not found at long-term exposures (7 or 10 days), except that TNMD expression was maintained at 50 ng/mL at a 7-day exposure to TGF-β3. Like genes, the protein expression levels of COL1, TNMD, and SCX were also induced in TGF-β3-treated TMSCs in a 3-day treatment, which were maintained for 10 days, as evidenced by immunofluorescence staining, immunohistochemistry and Western blot analyses. This study demonstrated that TMSCs in tenogenic stimulation with TGF-β3 have a high tenogenic differentiation potential.

Keyword

Transforming growth factor beta 3; Tonsil-derived mesenchymal stem cells; Tenogenesis

MeSH Terms

Blotting, Western
Collagen
Fluorescent Antibody Technique
Humans
Immunohistochemistry
Mesenchymal Stromal Cells*
Palatine Tonsil
Polymerase Chain Reaction
Stem Cells
Tonsillectomy
Transforming Growth Factor beta
Collagen
Transforming Growth Factor beta
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