Expression of OCT-4 and SOX-2 in Bone Marrow-Derived Human Mesenchymal Stem Cells during Osteogenic Differentiation

Igor Matic; Maja Antunovic; Sime Brkic; Pavle Josipovic; Katarina Caput Mihalic; Ivan Karlak; Alan Ivkovic; Inga Marijanovic

doi:10.3889/oamjms.2016.008

Authors

Igor Matic Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb
Maja Antunovic Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb
Sime Brkic Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb
Pavle Josipovic Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb
Katarina Caput Mihalic Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb
Ivan Karlak Department of Traumatology, University Hospital Sestre Milosrdnice, Zagreb
Alan Ivkovic Clinical Hospital Sveti Duh, Zagreb
Inga Marijanovic Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb

DOI:

https://doi.org/10.3889/oamjms.2016.008

Keywords:

pluripotency markers, human mesenchymal stem cells, osteodifferentiation

Abstract

AIM: Determine the levels of expression of pluripotency genes OCT-4 and SOX-2 before and after osteogenic differentiation of human mesenchymal stem cells (hMSCs).

METHODS: Human MSCs were derived from the bone marrow and differentiated into osteoblasts. The analyses were performed on days 0 and 14 of the cell culture. In vitro differentiation was evaluated due to bone markers â€“ alkaline phosphatase (AP) activity and the messenger RNA (mRNA) expression of AP and bone sialoprotein (BSP). The OCT-4 and SOX-2 expression was evaluated at mRNA level by real-time qPCR and at protein level by immunocytochemistry.

RESULTS: In vitro cultures on day 14 showed an increase in AP activity and upregulation of AP and BSP gene expression. OCT-4 and SOX-2 in undifferentiated hMSCs on day 0 is detectable and very low compared to tumor cell lines as a positive control. Immunocytochemistry detected OCT-4 in the cell nuclei prior (day 0) and post differentiation (day 14). On the same time points, cultures were negative for SOX-2 protein.

CONCLUSION: Messenger RNA for pluripotency markers OCT-4 and SOX-2 isolated from hMSCs was less present, while OCT-4 protein was detected in cell nuclei prior and post differentiation into osteoblast lineage.

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