2013)

2013). of Nestin was diminished in PB-MSCs by attenuating BMP signaling. The obtained results suggested that BMP signaling might have a regulatory function on the Nestin expression in mesenchymal stem cells. value?=?0.022 and 0.006 for N50 and N75; Fig. ?Fig.44 and Table ?Table1).1). The obtained findings suggested that the inhibition of BMP signaling by Noggin may support differentiation of PB-MSCs through down regulation of Nestin expression. Open in a separate window Fig. 3 Nestin expression analysis in PB-MSCs treated with different concentratons of Noggin (N50; 50?ng/ml; N75; 75?ng/ml and N100; 100?ng/ml). Untreated PB-MSCs were used as control. The X-axis and Y-axis represented cell cultures ID and Relative expression level of Nestin in treated PB-MSCs as compared with untreated cells (control), respectively. PB-MSCs treated with 50?ng/ml (N50) and 75?ng/ml (N75) Noggin were associated with a significant reduction in the Nestin expression Open in a separate window Fig. 4 Nestin expression level in six cultures of PB-MSCs after treatment with 50?ng/ml Noggin (value?=?0.022, students t-test). Numbers I, II, III, IV, V and VI represented the independent cell cultures of treated PBMSCs Table 1 The statistical analysis of expression data obtained from three independent PB-MSCs cultures treated with 75?ng/ml Noggin valueStandard Error *The result is significant at em p /em ? ?0.05 Expression of neuronal markers in noggin-treated PB-MSCs To examine whether Noggin can MX1013 modulate the neuronal differentiation of PB-MSCs, the expression of neuronal markers was studied using qPCR at three independent cell cultures treated with 75?ng/ml Noggin. The results showed that Nestin, beta tubulin III and MAP2 were decreased in PB-MSCs treated with 75?ng/ml Noggin (Fig. ?(Fig.55 and Table ?Table1).1). The NFM and NSE expression pattern showed the conflicting results in different cell cultures. The expressions of SLITRK4, MECP2 and GABRA3 were also evaluated in two Noggin-treated PB-MSCs cultures (75?ng/ml). Figure ?Figure66 showed that the treatment was accompanied by decreased expression of SLITRK4. In contrast, the GABRA3 and MECP2 expression levels increased in PB-MSCs following the treatment with Noggin. Open in a separate window Fig. 5 The expression of neuronal markers including Nestin, beta tubulin III, MAP2, NFM and NSE in PBMSCs cultures ( em N /em ?=?3). The X-axis and Y-axis represented neuronal markers and Relative expression level MX1013 of each marker in treated PB-MSCs as compared with untreated cells (control), respectively. The mean (M) and standard error (SE) of each neuronal marker were shown in Table ?Table1.1. Numbers I, II and III represented the independent cell cultures of treated PBMSCs. Untreated PB-MSCs were used as control Open in a separate window Fig. 6 Results of SLITRK4, GABRA3 and MECP2 expression analysis in two independent Noggin-treated PB-MSCs ( em N /em ?=?2). The X-axis and Y-axis represented cell cultures ID and relative expression level of neuron-specific genes in treated PB-MSCs as compared with untreated cells (control), respectively. The expression of GABRA3 and MECP2 increased in PBMSCs after treatment. In contrast to GABRA3 and MECP2, treatment with Noggin decreased SLITRK4 expression in PBMSCs as compared with untreated cells. Numbers I and II represented the independent cell cultures of treated PBMSCs. Untreated PB-MSCs were used as control Discussion In recent years, numerous signaling pathways were known to induce proliferation and differentiation of mesenchymal stem cells into adipocytes and osteocytes (James 2013; Longobardi et al. 2006). Different studies indicated that several members of the BMP signaling pathway involved in the differentiation of MSCs into osteocytes, adipocytes and chondrocytes (Kang et al. 2009; Cheng et al. 2001; Majumdar et al. 2001; Lou et al. 1999). Besides, it has been widely known that EGF and bFGF display a crucial role in promoting survival and proliferation of MSCs (Fan et al. 2007; Rabbit polyclonal to AKAP5 Solchaga et al. 2005; Benavente et al. 2003). Also, high levels of bFGF expression were associated with MX1013 normal development of the nervous system (Dono et al. 1998). In the present study, the MX1013 floating cells with round shape were observed 2?days after the addition of bFGF and EGF. They were reminiscent of neural stem cells (NSCs). These observations provided evidence suggesting an important role for bFGF and EGF in stimulation of PB-MSCs proliferation. Co-culture of neural cells and MSCs derived from bone marrow has been demonstrated to be accompanied by enhanced Nestin expression (Aizman et al. 2013). Some studies indicated that MSCs can have the stimulating effects on differentiation of neural precursors (Robinson et al. 2011; Bai et al. 2007; Kang et al. 2003). Furthermore, it has been revealed that culture of MSCs on plates coated with Extracellular matrix (ECM) could provide more.