MicroRNA Replacing Oncogenic Klf4 and c-Myc for Generating iPS Cells via Cationized Pleurotus eryngii Polysaccharide-based Nanotransfection.
Induced pluripotent stem cells (iPSCs), resulting from the forced expression of cocktails out of transcription factors, such as Oct4, Sox2, Klf4, and c-Myc (OSKM), has shown tremendous potential in regenerative medicine. Although rapid progress has been made recently in the generation of iPSCs, the safety and efficiency remain key issues for further application. In this work, microRNA 302-367 was employed to substitute the oncogenic Klf4 and c-Myc in the OSKM combination as a safer strategy for successful iPSCs generation. The negatively charged plasmid mixture (encoding Oct4, Sox2, miR302-367) and the positively charged cationized Pleurotus eryngii polysaccharide (CPEPS) self-assembled into nanosized particles, named as CPEPS-OS-miR nanoparticles, which were applied to human umbilical cord mesenchymal stem cells for iPSCs generation after characterization of the physicochemical properties. The CPEPS-OS-miR nanoparticles possessed spherical shape, ultrasmall particle size, and positive surface charge. Importantly, the combination of plasmids Oct4, Sox2, and miR302-367 could not only minimize genetic modification but also show a more than 50 times higher reprogramming efficiency (0.044%) than any other single or possible double combinations of these factors (Oct4, Sox2, miR302-367). Altogether, the current study offers a simple, safe, and effective self-assembly approach for generating clinically applicable iPSCs.
Department of Pharmaceutics, School of Pharmacy, and Center for Drug/Gene Delivery and Tissue Engineering, Jiangsu University , Zhenjiang 212001, People's Republic of China.
Wenwen
W
Deng
Department of Pharmaceutics, School of Pharmacy, and Center for Drug/Gene Delivery and Tissue Engineering, Jiangsu University , Zhenjiang 212001, People's Republic of China.
Xia
X
Cao
Department of Pharmaceutics, School of Pharmacy, and Center for Drug/Gene Delivery and Tissue Engineering, Jiangsu University , Zhenjiang 212001, People's Republic of China.
Jingjing
J
Chen
Center for Drug/Gene Delivery and Tissue Engineering, and School of Medical Science and Laboratory Medicine, Jiangsu University , Zhenjiang 212001, People's Republic of China.
Zhijian
Z
Zhang
School of Life Science & Technology, China Pharmaceutical University , Nanjing 210009, People's Republic of China.
Qingtong
Q
Yu
Department of Pharmaceutics, School of Pharmacy, and Center for Drug/Gene Delivery and Tissue Engineering, Jiangsu University , Zhenjiang 212001, People's Republic of China.
Yan
Y
Wang
Center for Drug/Gene Delivery and Tissue Engineering, and School of Medical Science and Laboratory Medicine, Jiangsu University , Zhenjiang 212001, People's Republic of China.
Genbao
G
Shao
Department of Pharmaceutics, School of Pharmacy, and Center for Drug/Gene Delivery and Tissue Engineering, Jiangsu University , Zhenjiang 212001, People's Republic of China.
Jie
J
Zhou
School of Life Science & Technology, China Pharmaceutical University , Nanjing 210009, People's Republic of China.
Xiangdong
X
Gao
Department of Pharmaceutics, School of Pharmacy, and Center for Drug/Gene Delivery and Tissue Engineering, Jiangsu University , Zhenjiang 212001, People's Republic of China.
Jiangnan
J
Yu
Department of Pharmaceutics, School of Pharmacy, and Center for Drug/Gene Delivery and Tissue Engineering, Jiangsu University , Zhenjiang 212001, People's Republic of China.
Ximing
X
Xu
D000818:Animals; D002412:Cations; D016923:Cell Death; D002454:Cell Differentiation; D065150:Cellular Reprogramming / Q000187:drug effects; D024202:Electrophoretic Mobility Shift Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism*; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D036103:Nanotechnology / Q000379:methods*; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D020076:Pleurotus / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry*; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D002412:Cations; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D035683:MicroRNAs; D050814:Octamer Transcription Factor-3; D011134:Polysaccharides; D016271:Proto-Oncogene Proteins c-myc; D055748:SOXB1 Transcription Factors
induced pluripotent stem cells; microRNAs; nanoparticles; polysaccharides; self-assembly