Basic & Clinical Medicine ›› 2011, Vol. 31 ›› Issue (6): 630-635.

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Culture of mouse cortex neural stem cell and miRNAs’ influence on NSCs’ differentiation ability

Xiang-yu ZHAO1,Peng-cheng SHU2,Jing ZHANG2,Bin YIN3,Xiao-zhong PENG1   

  1. 1. (National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS&PUMC
    3. National Laboratory of Medicine Molecular Biology, IBMS, CAMS,School of Basic Medicine PUMC
  • Received:2011-04-06 Revised:2011-04-11 Online:2011-06-05 Published:2011-06-06
  • Contact: Xiao-zhong PENG

Abstract: Objective To establish the mouse cortex neural stem cells (NSCs) culture technology platform in vitro and to explore some miRNA’ influence on neural stem cells’ differentiation ability. Methods The NSCs were prepared after isolation from the E14 embryonic mouse cortex and cultured in proliferation medium, the cells were then cultured in the differentiation medium with the immunostaining method to detect whether they can differentiate into neurons and astrocytes normally. Meanwhile Realtime-PCR was used to detect the expression level of several miRNAs between NSCs and the differentiated cells,miRNAs or the inhibitor were then transfected into the NSCs by the high efficiency of lipid reagent, western blot was used to detect the miRNAs or the inhibitor’s influence on the differentiation ability of NSCs. Results Establish the model of mouse neural stem cell culture platform successfully, detect that a set of miRNAs expressed differently between the NSCs and differentiated cells, Western-Blot showed that the antisense of miRNA-124 could reduce the expression of neural specific marker protein β-tubulin III obviously (P<0.01), while overexpression of miR-137,128 could upregulate β-tubulin III somehow. Conclusion the lipid reagent can transfect the miRNAs into the NSCs efficiently and miR-124,137,128 probably play an important role in the differentiation of NSCs.

Key words: Neural stem cells, Neurons, Astrocytes, miRNA, Transfection, Differentiation

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