In this study, the role of hydrostatic pressure on staurosporine-induced neural differentiation in mouse bone marrow mesenchymal stem cells were investigated. The cells were cultured in treatment medium containing 100 nM of staurosporine for 4 hours; then the cells were affected by hydrostatic pressure (0, 25,50, 100 mmHg). The percentage of cell viability by trypan blue staining and the percentage of cell death by Hoechst/PI differential staining were assessed. We obtained the total neurite length. Expression of β-tubulin III and GFAP (Glial fi brillary acidic protein) proteins were also analyzed by immunocytochemistry. The percentage of cell viability in treatments decreased relative to the increase in hydrostatic pressure and time (p < 0.05). The results revealed that total neurite length increased in cells treated with 25 mmHg hydrostatic pressure and decreased in those treated with 100 mmHg hydrostatic pressure (p < 0.05). Low hydrostatic pressure as a mechanical force can improve neural differentiation in mMSCs (mouse mesenchymal stem cells) (Tab. 3, Fig. 7, Ref. 30). Text in PDF www.elis.sk.

Background: Human umbilical cord mesenchymal stem cells (hUMSCs) have been considered to repair damaged tissues and cells. This study aims to investigate the differentiation efficiency affected by Schwann cells (SCs) and laminin and also compare them to other strategies using chemicals or growth factors. Methods: SCs and hUMSCs were separated from dorsal root ganglion of rats and newborn human umbilical cords (hUCs), respectively, and then cultured. The marker expressions of mesenchymal stem cells (MSCs), hematopoietic and endothelial for hUMSCs were confirmed by flow cytometry. The hUMSCs were cultured in four groups: 1) control, 2) co-culture with SCs (C), 3) laminin (L), and 4) co-culture with SCs treated by laminin (CL). The expression of protein and gene-related differentiation NSE, MAP2 and β-tubulin were examined by real-time polymerase chain reaction (PCR) and immunocytochemistry after 12 days. Results: The flow cytometry analysis revealed high expression of mesenchymal and low expression of hematopoietic and endothelial markers, where the SCs expressed S100 at a high level (97.4% ± 2.25). The expression of NSE, MAP2 and β-tubulin increased significantly in the C, L and CL groups compared to the control group (P < 0.001), where the CL group had the highest expression among the groups [7.59 ± 0.126, 7.87 ± 0.191, 6.36 ± 0.420, respectively, (P < 0.01)]. Also, the expression of neural proteins was significantly increased in tested groups in comparison to the control group. Conclusion:Combined laminin and SCs co-culturing with hUMCSs could be the most effective strategy for neural differentiation.