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


CONCLUSIONS
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.
Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia.
Igor
I
Matic
Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia.
Maja
M
Antunovic
Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia.
Sime
S
Brkic
Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia.
Pavle
P
Josipovic
Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia.
Katarina Caput
KC
Mihalic
Department of Traumatology, University Hospital Sestre Milosrdnice, Zagreb, Croatia.
Ivan
I
Karlak
Clinical Hospital Sveti Duh, Zagreb, Croatia.
Alan
A
Ivkovic
Division of Biology, Department of Molecular Biology, Faculty of Science, University of Zagreb, Zagreb, Croatia.
Inga
I
Marijanovic
human mesenchymal stem cells; osteodifferentiation; pluripotency markers