A Novel Protocol to Differentiate Induced Pluripotent Stem Cells by Neuronal microRNAs to Provide a Suitable Cellular Model.
作者： Mehrak Zare , Masoud Soleimani , Abolfazl Akbarzadeh , Behnaz Bakhshandeh , Seyed Hamid Aghaee-Bakhtiari
DOI: 10.1111/CBDD.12485
关键词: Downregulation and upregulation 、 Bioinformatics 、 microRNA 、 Cellular model 、 Immunocytochemistry 、 Biology 、 Gene 、 Epigenetics 、 Induced pluripotent stem cell 、 Enolase 、 Cell biology
摘要: Neurodegenerative diseases are one of the most challenging subjects in medicine. Investigation their underlying genetic or epigenetic factors is hampered by lack suitable models. Patient-specific induced pluripotent stem cells (iPS cells) represent a valuable approach to provide proper model for poorly understood mechanisms neuronal and related drug screenings. miR-124 miR-128 two brain-enriched miRNAs with different time-points expression during development. Herein, we transduced human iPS harboring lentiviruses sequentially. The plasmids contained GFP puromycin antibiotic-resistant genes easier selection identification. Morphological assessment immunocytochemistry (overexpressions beta-tubulin neuron-specific enolase) confirmed that hiPS similar characteristics those mature neurons. In addition, upregulation enolase, beta-tubulin, Map2, GFAP, BDNF was detected quantitative real-time PCR. conclusion, it seems our novel protocol remarks combinatorial effect on neural differentiation absence any extrinsic factor. Moreover, such cellular models could be used personalized screening applied more effective therapies.
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