C/EBP-α and C/EBP-β–mediated adipogenesis of human mesenchymal stem cells (hMSCs) using PLGA nanoparticles complexed with poly(ethyleneimmine)
| Autor: | Hyun-Jin Do, Jae Hwan Kim, Ji Sun Park, Han Na Yang, Keun-Hong Park, Dae Gyun Woo, Hye-Young Lim, Su Yeon Jeon |
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| Rok vydání: | 2011 |
| Předmět: |
Materials science
Cellular differentiation Blotting Western Biophysics Mice Nude Bioengineering macromolecular substances Green fluorescent protein Biomaterials Mice chemistry.chemical_compound Polylactic Acid-Polyglycolic Acid Copolymer CCAAT-Enhancer-Binding Protein-alpha Animals Humans Polyethyleneimine Lactic Acid DNA Primers Mice Inbred BALB C Base Sequence Reverse Transcriptase Polymerase Chain Reaction CCAAT-Enhancer-Binding Protein-beta Mesenchymal stem cell technology industry and agriculture Cell Differentiation Mesenchymal Stem Cells Transfection Molecular biology In vitro Blot PLGA Adipose Tissue chemistry Mechanics of Materials Adipogenesis Ceramics and Composites Nanoparticles Female Polyglycolic Acid |
| Zdroj: | Biomaterials. 32:5924-5933 |
| ISSN: | 0142-9612 |
| DOI: | 10.1016/j.biomaterials.2011.04.072 |
| Popis: | In this study, to drive efficient adipogenic differentiation, the adipogenic transcription factors C/EBP-α and C/EBP-β fused to green fluorescent protein (GFP) or red fluorescent protein (RFP) were complexed with poly-ethyleneimine (PEI) coupled with biodegradable PLGA nanospheres and delivered to human mesenchymal stem cell (hMSC). FACS analysis revealed that the transfection efficiency of C/EBP-α, C/EBP-β, or both genes complexed with PEI-coated PLGA nanospheres was 12.59%, 21.74%, and 28.96% of hMSCs. Expression and localization of C/EBP-α and C/EBP-β were confirmed by Western blotting and confocal laser microscopy. Overexpression of exogenous C/EBP-α and C/EBP-β significantly elevated adipogenic differentiation processes as indicated by RT-PCR, real-time PCR, Western blotting, histology, and immunofluorescence microscopy. During adipogenesis, PEI-coupled PLGA nanospheres complexed with C/EBP-α and C/EBP-β greatly increased the adipogenic capability of in vitro cultured cells, as well of in vivo transplanted cells. The expression of genes and proteins specific to adipogenic differentiation in hMSCs was significantly elevated compared to the controls. |
| Databáze: | OpenAIRE |
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