Gene Transfection toward Spheroid Cells on Micropatterned Culture Plates for Genetically-modified Cell Transplantation.
To improve the therapeutic effectiveness of cell transplantation, a transplantation system of genetically modified, injectable spheroids was developed. The cell spheroids are prepared in a culture system on micropatterned plates coated with a thermosensitive polymer. A number of spheroids are formed on the plates, corresponding to the cell adhesion areas of 100 µm diameter that are regularly arrayed in a two-dimensional manner, surrounded by non-adhesive areas that are coated by a polyethylene glycol (PEG) matrix. The spheroids can be easily recovered as a liquid suspension by lowering the temperature of the plates, and their structure is well maintained by passing them through injection needles with a sufficiently large caliber (over 27 G). Genetic modification is achieved by gene transfection using the original non-viral gene carrier, polyplex nanomicelle, which is capable of introducing genes into cells without disrupting the spheroid structure. For primary hepatocyte spheroids transfected with a luciferase-expressing gene, the luciferase is sustainably obtained in transplanted animals, along with preserved hepatocyte function, as indicated by albumin expression. This system can be applied to a variety of cell types including mesenchymal stem cells.
Graduate School of Medicine, Laboratory of Clinical Biotechnology, The University of Tokyo; itaka-ort@umin.net.
Keiji
K
Itaka
Graduate School of Medicine, Laboratory of Clinical Biotechnology, The University of Tokyo.
Satoshi
S
Uchida
Graduate School of Medicine, Laboratory of Clinical Biotechnology, The University of Tokyo.
Akitsugu
A
Matsui
Graduate School of Medicine, Laboratory of Clinical Biotechnology, The University of Tokyo.
Kayoko
K
Yanagihara
Graduate School of Medicine, Laboratory of Clinical Biotechnology, The University of Tokyo.
Masaru
M
Ikegami
Graduate School of Engineering, Department of Materials Engineering, The University of Tokyo.
Taisuke
T
Endo
Graduate School of Engineering, Department of Bioengineering, The University of Tokyo.
Takehiko
T
Ishii
Graduate School of Engineering, Department of Materials Engineering, The University of Tokyo.
Kazunori
K
Kataoka
D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D017690:Cell Transplantation / Q000379:methods*; D015870:Gene Expression; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology* / Q000637:transplantation; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology; D011092:Polyethylene Glycols / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D018874:Spheroids, Cellular / Q000166:cytology / Q000502:physiology* / Q000637:transplantation; D014162:Transfection / Q000379:methods*; D019076:Transgenes
D011092:Polyethylene Glycols; D008156:Luciferases