NGF combined with b-FGF induces differentiation of embryonic rat septum derived neural stem cells into neurons

ZHU Qing; CHEN Yan; LONG Da-hong; YANG Dan-di; XU Li-ping; WANG Di

doi:10.3969/j.issn.1001-9448.2019.05.006

Abstract：Objective To investigate the effect of NGF combined with b-FGF on the differentiation of embryonic rat septum derived neural stem cells into neurons.Methods Embryonic rat septum derived neural stem cells were cultured in vitro. The differentiation capacity, and the expression of nestin, GFAP, β-Tubulin Ⅲ were detected by cell IF staining. The neural spheres were divided into control group, NGF group (NGF 50 ng/mL), NGF combined with b-FGF (NGF 50 ng/mL, and gradually remove the b-FGF) group. The ratio of β-Tubulin Ⅲ positive cells were detected by IF staining. Results The neural stem cells derived from embryonic rats septum can be obtained a large number of nestin positive neurospheres through serum-free culture containing EGF and b-FGF. When removing mitogenic factors EGF and b-FGF, the stem cells could be naturally differentiated into GFAP and β-Tubulin Ⅲ positive cells. In the control group, NGF group and NGF combined with b-FGF group, the ratio of β-Tubulin Ⅲ positive cells were (21.80±2.81) ％, (38.04±3.86) ％ and (50.01％±7.65) ％, respectively. The neuronal differentiation rate in the combined group was significantly higher than NGF group (P＜0.05), and they were both significantly higher than that in control group (P＜0.05). Conclusion Nestin positive NSC can be isolated and cultured from the septum of embryonic rats, these neurospheres can be naturally differentiated into GFAP and β-Tubulin Ⅲ positive cells. NGF can induce differentiation of neural stem cells from septum origin into neurons in vitro, when gradually removing the b-FGF, the differentiation rate of neurons increases.