Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells

Abstract

Small molecules, growth factors, and cytokines have been used to induce differentiation of stem cells into different lineages. Similarly, demethylating agents can trigger differentiation in adult stem cells. Here, we investigated the in vitro differentiation of rat bone marrow mesenchymal stem cells (MSCs) into cardiomyocytes by a demethylating agent, zebularine, as well as neuronal-like cells by β-mercaptoethanol in a growth factor or cytokines-free media. Isolated bone marrow-derived MSCs cultured in Dulbecco’s Modified Eagle’s Medium exhibited a fibroblast-like morphology. These cells expressed positive markers for CD29, CD44, and CD117 and were negative for CD34 and CD45. After treatment with 1 μM zebularine for 24 hours, the MSCs formed myotube-like structures after 10 days in culture. Expression of cardiac-specific genes showed that treated MSCs expressed significantly higher levels of cardiac troponin-T, Nkx2.5, and GATA-4 compared with untreated cells. Immunocytochemical analysis showed that differentiated cells also expressed cardiac proteins, GATA-4, Nkx 2.5, and cardiac troponin-T. For neuronal differentiation, MSCs were treated with 1 and 10 mM β-mercaptoethanol overnight for 3 hours in complete and serum-free Dulbecco’s Modified Eagle’s Medium, respectively. Following overnight treatment, neuron-like cells with axonal and dendritic-like projections originating from the cell body toward the neighboring cells were observed in the culture. The mRNA expression of neuronal-specific markers, Map2, Nefl, Tau, and Nestin, was significantly higher, indicating that the treated cells differentiated into neuronal-like cells. Immunostaining showed that differentiated cells were positive for the neuronal markers Flk, Nef, Nestin, and β-tubulin.

Zebularine Promotes Hepatic Differentiation of Rabbit Bone Marrow Mesenchymal Stem Cells by Interfering with p38 MAPK Signaling

“…However, a number of studies have investigated the effect of zebularine on hepatic tumors and cardiomyocyte differentiation. It has been reported that zebularine inhibits liver tumor cell proliferation, induces apoptosis, and promotes their maturation and differentiation in a concentration-, time-, and dose-dependent manner [ 26 , 27 ]. In another study, it has been demonstrated that different doses of zebularine have opposite roles in immunogenicity [ 28 ].…”

Differentiation of mesenchymal stem cells from human amniotic fluid to cardiomyocyte-like cells

“…Previous studies have shown that MSCs from bone marrow can improve cardiac function in myocardial infarction ( 21 , 22 ) and cardiogenic differentiation from bone marrow MSCs has long been investigated ( 23 , 24 ). However, the process of using MSCs obtained from this source has displayed certain limitations including the invasive technique needed to obtain MSCs from bone marrow, the decreased capacity of these cells that correlate with the increased age of the donor ( 25 ) and the obstacle of cell transplantation therapy by immunological rejection ( 26 ).…”

Neuron-Specific Fluorescence Reporter-Based Live Cell Tracing for Transdifferentiation of Mesenchymal Stem Cells into Neurons by Chemical Compound

“…Many studies have reported on transdifferentiation of MSCs into neuronal lineage by treatment of chemicals such as β -mercaptoethanol (BME)/dimethyl sulfoxide (DMSO)/butylated hydroxyanisole or neurotrophic factors or by their coculture with neural or glial cells [ 31 – 37 ]. More recently, the BME-treated MSCs showed neuron-like features, expressing high level of neural-specific markers (Map2, Nefl, Tau, and nestin) [ 38 ]. However, previous neuronal induction protocol using chemicals showed relatively low efficiency and toxic effect to cultured cells [ 39 , 40 ].…”