Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) could be induced to differentiate into neuron-like cells through a variety of inducers in vitro. Different inducers of cell differentiation rate, cell viability, as well as the rate of nestin-positive cells staining are different.
OBJECTIVE: To explore a better inducer by comparing results of BMSCs differentiation into neuron-like cells using multiple inducers.
METHODS: The healthy adult bone marrow was taken in the sterile laminar flow room. Lymphocyte separating medium was added for centrifugation, and the albuginea was obtained. Cells were placed in medium containing basic fibroblast growth factor (bFGF), epidermal growth factor, DMEM and 10% fetal bovine serum in 5% CO₂ incubator at 37 ℃. The third passage of cells were divided into β-mercaptoethanol + dimethyl sulfoxide group, brain-derived neurotrophic factor (BDNF) + retinoic acid group, glial cell line-derived neurotrophic factor (GDNF) + retinoic acid group and bFGF group. Inducers were added separately. Cell morphology was observed. Cell viability was measured. Nerve cell markers were detected using immunohistochemistry.
RESULTS AND CONCLUSION: Cell viability was compared in each group using trypan blue staining. Cell viability was greatest in the GDNF + retinoic acid group (P < 0.05), but lowest in the β-mercaptoethanol + dimethyl sulfoxide group (P < 0.05). No significant difference in cell viability was determined between the BDNF + retinoic acid group and bFGF group (P > 0.05). Positive rate of nerve cell markers was greater in the GDNF + retinoic acid group compared with β-mercaptoethanol + dimethyl sulfoxide group, BDNF + retinoic acid group and bFGF group (P < 0.05). These suggest that adult BMSCS could differentiate into neuron-like cells. GDNF + retinoic acid was the optimal induction which showed higher living cell energy and positive immunoreactivity to nerve markers (P < 0.05).