The aim of this study was to investigate the effect of gap junction intercellular communication on the valproic acid-induced differentiation of adipose tissue-derived stem cells (ADSCs) into Schwann cells. Methods: ADSCs from rats were isolated and cultured. At passage 3, the cells were inoculated with medium containing no valproic acid (group A), 1.0 mmol/L valproic acid (group B), or 1.0 mmol/L valproic acid + 2.5 μmol/L oleamide (group C). Changes in cell morphology were observed by microscopy. Immunocytochemical staining and real-time quantitative PCR were employed to assess the protein expression and mRNA transcription levels of S100, neuron-specific enolase (NSE), Nestin, and Connexin-43 (Cx43). Results: Compared to group A, groups B and C showed a significant increase in the protein expression and mRNA transcription levels of S100, NSE, and Nestin (P<0.01), and group C showed lower levels than group B (P<0.01). Groups B and C also showed significantly higher levels of protein expression and mRNA transcription of Cx43 than group A, and there was no significant difference between group B and group C (P>0.05). Conclusion: Cx43-mediated gap junction intercellular communication may serve an important role in the valproic acid-induced differentiation of ADSCs into Schwann cells.