Document Detail


Efficient preparation and labeling of human induced pluripotent stem cells by nanotechnology.
MedLine Citation:
PMID:  21499432     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Efficient preparation and labeling of human induced pluripotent stem (iPS) cells is a great challenge in stem cell research and development. With the aim of investigating the feasibility of using nanotechnology to enhance the preparation efficiency of iPS cells and to label iPS cells for long-term tracing and imaging, in this paper, four transcription factor genes, ie, Oct4, Sox2, LIN28, and Nanog, and packaging plasmids such as PSPAX2 and PMD2.G were cotransfected into 293T cells using Generation 5.0 polyamidoamine dendrimer-modified magnetic nanoparticles (dMNPs) as a delivery system. The resultant supernatant liquids were incubated with human fibroblast cells at 37°C for 21 days, then the embryonic stem (ES) cell-like clones were screened, cultured, and identified. Finally, the prepared iPS cells were labeled with fluorescent magnetic nanoparticles (FMNPs). The results showed that dMNPs can efficiently deliver all vectors into 293T cells. The resultant lentiviruses' titers were 10-fold more than those based on Lipofectamine™ 2000. Reverse transcription polymerase chain reaction analysis showed that four genes (Oct4, Sox2, LIN28, and Nanog) exhibited different expressions in iPS cells. Immunostaining analysis showed that specific surface markers of ES cells such as SSEA-3, SSEA-4, Tra-1-60, and Tra-1-81 were positive in iPS cells, and the terotomas were formed in NOD-SCID mice that were implanted with iPS cells. Red fluorescent signals could be observed in iPS cells labeled with FMNPs by fluorescent microscopy, and the magnetic signals were detected in labeled iPS cells by magnetic resonance imaging. In conclusion, human iPS cells can be efficiently generated using polyamidoamine dMNPs and lentivirus and labeled with FMNPs for long-term observation and tracking, which has great potential application in the research and development of stem cells in the near future.
Authors:
Jing Ruan; Jie Shen; Zheng Wang; Jiajia Ji; Hua Song; Kan Wang; Bin Liu; Jinhui Li; Daxiang Cui
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2011-02-21
Journal Detail:
Title:  International journal of nanomedicine     Volume:  6     ISSN:  1178-2013     ISO Abbreviation:  Int J Nanomedicine     Publication Date:  2011  
Date Detail:
Created Date:  2011-04-18     Completed Date:  2011-08-24     Revised Date:  2013-06-30    
Medline Journal Info:
Nlm Unique ID:  101263847     Medline TA:  Int J Nanomedicine     Country:  New Zealand    
Other Details:
Languages:  eng     Pagination:  425-35     Citation Subset:  IM    
Affiliation:
Department of Bio-Nano Science and Engineering, Key Laboratory for Thin Film and Microfabrication Technology of the Ministry of Education, National Key Laboratory of Micro/Nano Fabrication Technology, Research Institute of Micro/Nano Science and Technology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Cell Separation
Cells, Cultured
DNA Primers / genetics
Dendrimers / chemistry
Flow Cytometry
Fluorescent Dyes / administration & dosage
Gene Expression
Genetic Vectors
Humans
Induced Pluripotent Stem Cells / cytology*,  metabolism,  transplantation
Karyotyping
Lentivirus / genetics
Magnetic Resonance Imaging
Magnetite Nanoparticles / administration & dosage,  chemistry
Mice
Mice, Inbred NOD
Mice, SCID
Microscopy, Fluorescence
Nanomedicine
Nanotechnology
Transcription Factors / genetics
Transfection
Transplantation, Heterologous
Chemical
Reg. No./Substance:
0/DNA Primers; 0/Dendrimers; 0/Fluorescent Dyes; 0/Magnetite Nanoparticles; 0/PAMAM Starburst; 0/Transcription Factors
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