, ,
Volume 12, Issue 3 (11-2012)
Abstract
Umbilical cord (UC) is an important source of multipotential mesenchymal stem cells (MSCs). MSCs can be differentiated into different types of cells if they cultured under specific conditions. It has been proved that vitamin D3 can cause differentiation of stem cells in to osteoblast. Also, it has been observed that bee venom (BV) is effective in differentiation of cancerous cells. In this study differentiating potential of BV and vitamin D3 on MSCs to osteoblast was examined. Furthermore, our hypothesis was that BV could increase differentiating potential of vitamin D3. The cells obtained from UC tissues of 10-12 mouse embryos which were digested enzymatically and suspended in DMEM medium. For approving of stem cells, embryonic Oct4 marker was checked and the mesenchymal character of these cells was proven by surface markers including CD73, CD29, CD44. The flowcytometric analysis revealed high levels of these markers. After the second passage, in order to induce osteogenic differentiation, cells were cultured for 21days in DMEM medium containing different concentrations of BV, vitamin D3 separately and BV/vitamin D3 together. At first, cytotoxic effects of BV on MSCs were tested by MTT assay, which were shown that BV inhibited MSCs growth at higher concentrations than 6&mug/ml. Following the treatment, calcium’s level in the cells was determined by Alizarin red staining. Also, as an osteogenic marker, alkalin phosphatase level was measured in treated and non-treated cells. By Alizarin red staining and alkalin phosphatase assay, we found that BV With non-toxic concentrations (2µgr/ml, 4µgr/ml and 6µgr/ml) can cause a few osteogenic differentiation of MSCs. Whereas, usage of BV/vitamin D3 together, caused increasing the differential effects of vitamin D3. In conclusion we suggest that if MSCs were treated with bee venom and vitamin 3 at the same time, these components able to differentiate MSCs to osteoblast, therefore, they could be useful in cell therapy.
