Sodium Butyrate Pre-Treatment Enhances Hepatic Differentiation of Bone Marrow Mesenchymal Stem Cells Under Liver-Specific Factors Induction In Vitro

Abstract

Aims: It is crucial to establish an effective method to improve mesenchymal stem cells (MSCs) hepatic differentiation in mesenchymal stem cells transplantation. Herein, we aim to explore whether sodium butyrate (NaB) enhances hepatic differentiation of bone marrow mesenchymal stem cells (BM-MSCs) under liver-specific factors induction in vitro. Methods: We isolated BM-MSCs from the mononuclear cell fraction of rabbit bone marrow samples, and identified these cells by immunophenotype analysis. We then investigated the effects of different concentrations and different induced conditions. The histone deacetylase inhibitor NaB induced hepatic differentiation of BM-MSCs under liver-specific factors induction in vitro. BM-MSCs cultured in basic medium without the differentiation stimuli were set as the control. Morphological features, liver-specific gene and protein expression, and functional analyses were assessed to evaluate hepatic differentiation of BM-MSCs. Results: Our results showed that the persistence of NaB inhibited the expression of liver-specific protein expression in a dose-dependent manner. When NaB was continuously induced, all the four concentrations used were not conducive to induce hepatocyte transformation, but the low concentrations (0.5 and 1.0 mM) were relatively better. We found that the induction efficiency of NaB 24h pre-treatment was higher than that of NaB continuous intervention for 0.5 mM or 1.0 mM conditions, and higher than liver-specific factors such as hepatocyte growth factor (AGF) and epidermal growth factor (EGF) without NaB. Conclusion: Continuous NaB treatment can inhibit BMMSCs proliferation with concentration-dependence in a certain concentration range. 24h pre-treatment can enhance hepatic differentiation of BM-MSCs under liver-specific factors induction in vitro.