On-chip transfection of PC12 cells based on the rational understanding of the role of ECM molecules: efficient, non-viral transfection of PC12 cells using collagen IV.
Transfection microarrays (TMA) are important emerging tools for the study of genetic events in living cells in a high-throughput fashion and with significant material economy. However, the difficulty to transfect various relevant cell types on-chip hinders the use of TMAs. Herein we present the realization of a transfection microarray applicable to PC12 cells that heavily relies on the use of ECM molecules. Collagen IV and at a lesser extent laminin or collagen I, but not fibronectin or poly-l-lysine were found to significantly increase the solution-phase as well as on-chip transfection efficiency of PC12 cells. The highest transfection efficiency obtained was consistently above 60%. The observed correlations between the transfection efficiencies and the differential adhesion-induced events triggered by the studied ECMs provides the basis for the rationalization of the role of ECMs on the transfection process.
Uchimura
Eiichiro
E
Research Institute of Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Nakouji 3-11-46, Amagasaki, Hyogo 661-0974, Japan.
Yamada
Shigeru
S
Uebersax
Lorenz
L
Yoshikawa
Tomohiro
T
Matsumoto
Kayo
K
Kishi
Michiko
M
Funeriu
Daniel P
DP
Miyake
Masato
M
Miyake
Jun
J
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology; D024042:Collagen Type I / Q000502:physiology; D024141:Collagen Type IV / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000502:physiology*; D005353:Fibronectins / Q000502:physiology; D007797:Laminin / Q000502:physiology; D016716:PC12 Cells; D051381:Rats; D014162:Transfection / Q000379:methods*
D024042:Collagen Type I; D024141:Collagen Type IV; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D007797:Laminin; D002118:Calcium