Abstract:

BACKGROUND: Cell plant density is one of the factors that affect stem cell differentiation. To date, there is still lack of further study about the effect of cell plant density on bone marrow mesenchymal stem cells differentiation into neuronal-like cells.
OBJECTIVE: To investigate the effect of cell density on the differentiation of rat bone marrow mesenchymal stem cells into neuronal-like cells.
METHODS: Rat bone marrow mesenchymal stem cells were isolated by use of adherent culture method. Following four passages, cells were cultured in six-well plates at a density of 2×10²/cm², 2×10³/cm², and 4×10³/cm², 8×10³/cm², 2×10⁴/cm², 4×10⁴/cm². Cells in each group were differentiated into neuronal-like cells under the induction of basic fibroblast growth factor + epidermal growth factor + RA, and were identified by immunohistochemical staining method. The proportion of neuronal-like cells in each group was calculated, and the differentiation rate was compared.
RESULTS AND CONCLUSION: Neuronal-like cells were observed after adding inducer in bone marrow mesenchymal stem cells, and Nestin, neuron-specific enolase, glial fibrillary acidic protein staining was positive. Different cell plant densities induced different ratios of neuron-like cells. 8×10³/cm² plant density yielded highest proportion of neuronal-like cells and can survive as long as 7 days. The results demonstrated that the differentiation of bone mesenchymal stem cells into neuron-like cells was correlated with cell plant density. Too high or too low cell plant density was detrimental to differentiation.