Comparative analysis of mouse embryonic palatal mesenchymal cells isolated by two primary culture methods

Cell culture plays a vital role in mechanism research, as a supplement to experiments in vivo. At present, there are two main methods to obtain mouse embryonic palatal mesenchymal (MEPM) cells, while no systematic investigation about characteristics of cells using these two methods to get a clear application. In this study, using the traditional two-step primary culture method as the control group, we found that the MEPM cells of the simplified one-step method and the control group were both consistent with the surface markers of mesenchymal stem cells. The vimentin (mesenchymal marker) was positive, cytokeratin 14 (CK14, epithelium marker) was negative. In addition, along with the increasing passages, the cells cultured by two methods had similar changes with a decreasing proliferation activity, increasing apoptosis rate, declining migration, diminishing stemness, and obvious senescence. Moreover, the senescence of MEPM cells in the one-step group was lower than that in the control group. Our findings indicate that there are no significant differences between the two methods and one-step method can be a good method in investigating palatal development mechanisms by shortening the processing of the MEPM cell culture and may reduce the damage to the cells done by more enzymes and processing.