Bio treatment protects rat marrow-derived mesenchymal stem cell culture against the TNF-a induced apoptosis

ABSTRACT

This study is an attempt to examine the anti apoptotic effects of BIO on rat MSC culture. Rat marrow primary cell culture was established and exposure groups were defined; cultures with 0.01, 0.1, 1 micro M BIO. Cells cultured without BIO treatment were used as controls. During culture expantion, the average doubling time, as an index of the rate of cell growth, were determined and compared. To examine whether or not BIO is able to protect MSCs against apoptosis, the passaged-3 cells from each group were induced to undergo apoptosis with the addition of TNF-alpha [Tumor necrotic factor-alpha]. Three days after, the cultures were quantified in terms of the percentages of apoptotic cells using either the Tunnel or Annexin V staining method. Marrow cells cultivated with 0.1 and 1 micro M BIO appeared to expand at a significantly more rapid rate than the 0.01 micro M BIO and the control cultures [p < 0.05]. Tunnel staining indicated that in 1 micro M BIO-treated groups, there were lower percentages of apoptotic nuclei than in groups with other concentrations of BIO [p < 0.05]. The BIO protective effect appeared to be dose-dependent in that the cultures with high BIO content possessed less apoptotic nuclei. The results obtained by Annexin staining were in agreement with the results of Tunnel staining. The Annexin method additionally takes into account the early apoptotic cells which are not detectable by the Tunnel method. Taken together, it seems that cultivation with BIO could both increase the growth rate of marrow cells and protect MSCs against induced apoptosis