Dual delivery of siRNA and plasmid DNA using mesoporous silica nanoparticles to differentiate induced pluripotent stem cells into dopaminergic neurons

作者：Chang Jen Hsuan; Tsai Ping Hsing; Chen Wei; Chiou Shih Hwa^*; Mou Chung Yuan^*

来源：Journal of Materials Chemistry B, 2017, 5(16): 3012-3023.

DOI：10.1039/c7tb00351j

The cells of the central nervous system (CNS) show irreversible features after injury, and they could be re-established by stem cells. Induced pluripotent stem cells (iPSCs) may be generated from adult mammalian cells and gain pluripotency to differentiate into various lineages and functional cells. Non-viral carriers for delivering differentiation factors to transform iPSCs into neuronal cells are very desirable. In this study, mesoporous silica nanoparticles (MSNs) are used to co-deliver Nurr1 plasmid DNA (pNurr1) and Rex1 siRNA (siRex1) into iPSCs to achieve dopaminergic neuron differentiation. Sixty hours after treatment with siRex1 and pNurr1, the co-delivery of pNurr1 and siRex1 enhanced the Nurr1 gene expression three-fold, compared to the delivery of the plasmid pNurr1 only. The dopaminergic neuron-related protein level was identified using immunofluorescence staining and flow cytometry analysis, and there were 89.9% +/- 0.5% tyrosine hydroxylase-expressing cells and 88.5% +/- 2.0% dopamine transporter-expressing cells differentiated from iPSCs after transfection by MSNs. An ELISA was also used to determine the maturation of functional neurons, and there was 12.19 ng mL(-1) dopamine released from cells after transfection by MSNs. These results demonstrate that MSNs are good non-viral nanocarriers for dual delivery of pNurr1 and siRex1 to significantly enhance the generation of dopaminergic neurons from iPSCs.

• 出版日期2017-4-28