Effects of low-intensity pulsed ultrasound on cell viability, proliferation and neural differentiation of induced pluripotent stem cells-derived neural crest stem cells

Abstract

Low-intensity pulsed ultrasound (LIPUS) acting on induced pluripotent stem cells-derived neural crest stem cells (iPSCs-NCSCs) is considered a promising therapy to improve the efficacy of injured peripheral nerve regeneration. Effects of LIPUS on cell viability, proliferation and neural differentiation of iPSCs-NCSCs were examined respectively in this study. LIPUS at 500 mW cm(-2) enhanced the viability and proliferation of iPSCs-NCSCs after 2 days and, after 4 days, up-regulated gene and protein expressions of NF-M, Tuj1, S100β and GFAP in iPSCs-NCSCs whereas after 7 days expression of only NF-M, S100β and GFAP were up-regulated. LIPUS treatment at an appropriate intensity can, therefore, be an efficient and cost-effective method to enhance cell viability, proliferation and neural differentiation of iPSCs-NCSCs in vitro for peripheral nerve tissue engineering.

In vivo repair of rat transected sciatic nerve by low-intensity pulsed ultrasound and induced pluripotent stem cells-derived neural crest stem cells

“…However, the optimal therapeutic intensity of LIPUS acted on the nerve conduit in vivo is still unclear. Our previously study (Lv et al 2013) proved that LIPUS at 0.3 and 0.5 W cm -2 could enhance the viability and proliferation of iPSCsNCSCs. Tsuang et al (2011) also demonstrated that 0.3 W cm -2 LIPUS could promote Schwann cells proliferation and prevent cell death in a severely injured peripheral nerve.…”

“…The regenerated neurons also could secrete some growth factors such as neural growth factor (NGF) and brained-derived neural growth factor (BDNF), which were benefit for the reconstruction of synapses. In our previously study (Lv et al 2013), the gene and protein Fig. 7 Immunostaining of Tuj1 of sciatic nerve in the cross section of nerve conduit at 5-7 mm from the proximal end at 3 months post-implantation.…”

“…On one hand, iPSCs-NCSCs can differentiate into vascular cells and neurons, which could accelerate the vascular and nerve regeneration during the restore of injured nerve (Kane et al 2011). On the other hand, LIPUS could induce angiogenesis (Hanawa et al 2014) and promote neural differentiation (Lv et al 2013). In addition, LIPUS could speed up the exchanges of nutrients and toxic substances which are good for the formation of nerve filaments (Chen et al 2010).…”

“…iPSCsNCSCs were cultured as described previously (Supporting Information) (Wang et al 2011;Lv et al 2013). Nerve conduits were sterilized with 75 % (v/v) ethyl alcohol before use.…”

“…Tsuang et al (2011) revealed that the intervention of low-intensity pulsed ultrasound (LIPUS) could promote primary cultured Schwann cells proliferation and prevent cell death, especially in a severely injured condition. In our previous research, the effect of LIPUS on the induced pluripotent stem cell-derived neural crest stem cells (iPSCs-NCSCs) was tested in vitro and the results showed that LIPUS had a positive effect on the viability, proliferation and neural differentiation of iPSCs-NCSCs (Lv et al 2013). The efficient and cost-effective method of LIPUS acting on iPSCs-NCSCs expanded their use on neural differentiation in vivo.…”