Abstract
Human deciduous periodontal ligament stem cells have been introduced for as an easily accessible source of stem cells from dental origin. Although recent studies have revealed the ability of these stem cells in multipotential attribute, their efficiency of hepatic lineage differentiation has not been addressed so far. The aim of this study is to investigate hepatic lineage fate competence of periodontal ligament stem cells through direct media induction. Differentiation of periodontal ligament stem cells into hepatocyte-like cells was conducted by the exposure of two phase media induction. First phase was performed in the presence of hepatocyte growth factors to induce a definitive endoderm formation. In the subsequent phase, the cells were treated with oncostatin M and dexamethosone followed by insulin and transferrin to generate hepatocyte-like cells. Hepatic-related characters of the generated hepatocyte-like cells were determined at both mRNA and protein level followed by functional assays. Foremost changes observed in the generation of hepatocyte-like cells were the morphological features in which these cells were transformed from fibroblastic shape to polygonal shape. Temporal expression of hepatic markers ranging from early endodermal up to late markers were detected in the hepatocyte-like cells. Crucial hepatic markers such as glycogen storage, albumin, and urea secretion were also shown. These findings exhibited the ability of periodontal ligament stem cells of dental origin to be directed into hepatic lineage fate. These cells can be regarded as an alternative autologous source in the usage of stem cell-based treatment for liver diseases.
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Acknowledgments
The work is part of an ongoing project between Hygieia Innovation and Faculty of Dentistry, University Malaya, and supported by University of Malaya, High Impact Research-Ministry of Higher Education, Malaysia (UM.C/HIR/MOHE/DENT/02).
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Communicated by: Sven Thatje
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ESM 1
Supplementary Figure 1. Bona fide MSCs features in isolated PDLSCs. A) Morphologically PDLSCs culture appeared spindle shaped (Scale bar = 100μm). B and C) Growth curve and population doubling time from P1 to P5 of respective cell culture (Scale bar = 200μm). D) Formation of colonies shown for PDLSCs. E) Percentage of senescent cells in PDLSCs. F) The panels display representative of visual observation (Scale bar = 200μm) of adipocytes detected by Oil Red O staining, chondrocytes detected by Alcian Blue staining and osteocytes detected by Von Kossa staining. Representative six independent result. (TIF 1866 kb)
ESM 2
Supplementary Figure 2. Immunophenotyping of mesenchymal stem cells surface markers. Isolated PDLSCs shown to be positively stained the gold standard of mesenchymal stem cells surface markers and absence of hematopoetic surface markers. (TIF 857 kb)
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Vasanthan, P., Jayaraman, P., Kunasekaran, W. et al. Generation of functional hepatocyte-like cells from human deciduous periodontal ligament stem cells. Sci Nat 103, 62 (2016). https://doi.org/10.1007/s00114-016-1387-7
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DOI: https://doi.org/10.1007/s00114-016-1387-7