engleski

Expression of pluripotency markers during mesenchymal stem cell differentiation

The purpose of this study was to determine the levels of expression of pluripotency genes Oct4, Sox2 and Nanog during the osteogenic differentiation of multipotent mesenchymal stem cells. Mesenchymal stem cells were derived from the human bone marrow and differentiated into osteoblasts. The analyses were performed on days 0 and 14 of the cell culture. In vitro differentiation was evaluated by the analysis of bone markers – alkaline phosphatase activity and the mRNA expression of dentin matrix protein 1 (DMP1) and bone sialoprotein (BSP). The Oct4, Sox2 and Nanog 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 alkaline phosphatase activity as well as the upregulation of DMP1 and BSP. Analysis of Oct4, Sox2 and Nanog transcripts in undifferentiated hMSC on day 0 showed that their expression decreases together with the appearance and increase of bone markers. However, immunocytochemistry detected only Oct4 transcription factor and it was localized in cell nuclei, prior (day 0) and post differentiation (day 14). On the same time points, cultures were negative for SOX2 and NANOG proteins. Conclusion: From presented data we conclude that pluripotency markers Oct4, Sox2 and Nanog are present in mesenchymal stem cells at mRNA level and that their transcription is decreasing as the differentiation proceeds. However, mRNA levels do not reflect the protein levels suggesting the posttranscriptional regulation of protein expression.

Mesenchymal stem cells; Oct4; Sox2; Nanog

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