Surface functionalization of inorganic nano-crystals with fibronectin and E-cadherin chimera synergistically accelerates trans-gene delivery into embryonic stem cells.
Stem cells holding great promises in regenerative medicine have the potential to be differentiated to a specific cell type through genetic manipulation. However, conventional ways of gene transfer to such progenitor cells suffer from a number of disadvantages particularly involving safety and efficacy issues. Here, we report on the development of a bio-functionalized inorganic nano-carrier of DNA by embedding fibronectin and E-cadherin chimera on the carrier, leading to its high affinity interactions with embryonic stem cell surface and accelerated trans-gene delivery for subsequent expression. While only apatite nano-particles were very inefficient in transfecting embryonic stem cells, fibronectin-anchored particles and to a more significant extent, fibronectin and E-cadherin-Fc-associated particles dramatically enhanced trans-gene delivery with a value notably higher than that of commercially available lipofection system. The involvement of both cell surface integrin and E-cadherin in mediating intracellular localization of the hybrid carrier was verified by blocking integrin binding site with excess free fibronectin and up-regulating both integrin and E-cadherin through PKC activation. Thus, the new establishment of a bio-functional hybrid gene-carrier would promote and facilitate development of stem cell-based therapy in regenerative medicine.
Kutsuzawa
K
K
Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Chowdhury
E H
EH
Nagaoka
M
M
Maruyama
K
K
Akiyama
Y
Y
Akaike
T
T
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D015820:Cadherins / Q000737:chemistry*; D002460:Cell Line; D004247:DNA / Q000737:chemistry / Q000493:pharmacokinetics*; D004337:Drug Carriers / Q000737:chemistry; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D005353:Fibronectins / Q000737:chemistry*; D007287:Inorganic Chemicals / Q000737:chemistry; D008422:Materials Testing; D051379:Mice; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D020033:Protein Isoforms / Q000737:chemistry; D013499:Surface Properties; D014162:Transfection / Q000379:methods*
D001672:Biocompatible Materials; D015820:Cadherins; D004337:Drug Carriers; D005353:Fibronectins; D007287:Inorganic Chemicals; D020033:Protein Isoforms; D004247:DNA