title: Transfection efficiency for size-separated cells synchronized in cell cycle by microfluidic device.
abstract: Non-viral system generally demonstrates less efficacious in transgene delivery than viral system; however it represents a safer alternative to viral system. In this study, transfection efficiency for human hepatocellular liver carcinoma cells synchronized in cell cycle at G0/G1 phase, which was sorted in size with a microfluidic device based on hydrodynamic filtration, was investigated by using a reverse transfection method. The synchronized cells were recovered at the yield of 80% from the micro-channel, and green fluorescent protein gene encoding plasmid mixed with lipofectoamine was transfected. The transfection efficiency of the cells at G0/G1 phase was 1.8 times higher than non-synchronized cells. The manipulation of cell cycle status could increase transfection efficiency in non-viral system, indicating size-based cell cycle synchronization is a powerful tool as a noninvasive method for bioscience and biotechnology.
authors.affiliation: Biomaterials Center, National Institute for Materials Science, 1-2-1 Sengen, Tsukuba, Ibaraki 305-0047, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Migita
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Hanagata
authors.forename: Daiju
authors.initials: D
authors.lastname: Tsuya
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Yamazaki
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Sugimoto
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Ikoma
mesh_terms: D002448:Cell Adhesion; D002469:Cell Separation; D004867:Equipment Design; D016193:G1 Phase / Q000235:genetics*; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D057446:Hydrodynamics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D010957:Plasmids; D016192:Resting Phase, Cell Cycle / Q000235:genetics*; D014162:Transfection / Q000379:methods*; D019076:Transgenes
chemical_list: D049452:Green Fluorescent Proteins