An efficient surface modification using 2-methacryloyloxyethyl phosphorylcholine to control cell attachment via photochemical reaction in a microchannel.
This report describes a direct approach for cell micropatterning in a closed glass microchannel. To control the cell adhesiveness inside the microchannel, the application of an external stimulus such as ultraviolet (UV) was indispensible. This technique focused on the use of a modified 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, which is known to be a non-biofouling compound that is a photocleavable linker (PL), to localize cells via connection to an amino-terminated silanized surface. Using UV light illumination, the MPC polymer was selectively eliminated by photochemical reaction that controlled the cell attachment inside the microchannel. For suitable cell micropatterning in a microchannel, the optimal UV illumination time and concentration for cell suspension were investigated. After selective removal of the MPC polymer through the photomask, MC-3T3 E1 cells and vascular endothelial cells (ECs) were localized only to the UV-exposed area. In addition, the stability of patterned ECs was also confirmed by culturing for 2 weeks in a microchannel under flow conditions. Furthermore, we employed two different types of cells inside the same microchannel through multiple removal of the MPC polymer. ECs and Piccells were localized in both the upper and down streams of the microchannel, respectively. When the ECs were stimulated by adenosine triphosphate (ATP), NO was secreted from the ECs and could be detected by fluorescence resonance energy transfer (FRET) in Piccells, which is a cell-based NO indicator. This technique can be a powerful tool for analyzing cell interaction research.
Jang
Kihoon
K
Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Sato
Kae
K
Tanaka
Yo
Y
Xu
Yan
Y
Sato
Moritoshi
M
Nakajima
Takahiro
T
Mawatari
Kazuma
K
Konno
Tomohiro
T
Ishihara
Kazuhiko
K
Kitamori
Takehiko
T
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002448:Cell Adhesion / Q000528:radiation effects; D002460:Cell Line; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism*; D008689:Methacrylates / Q000737:chemistry*; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D009569:Nitric Oxide / Q000032:analysis / Q000378:metabolism; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000737:chemistry; D055668:Photochemical Processes; D013499:Surface Properties / Q000528:radiation effects; D014466:Ultraviolet Rays
D008689:Methacrylates; D010767:Phosphorylcholine; D009569:Nitric Oxide; C070638:2-methacryloyloxyethyl phosphorylcholine; D000255:Adenosine Triphosphate