Cell patterning by micro-pattern projection of UV light through photoinduced enhancement of cell adhesion (PIECA).
Here we describe a protocol to prepare cell patterns by immobilizing anchor-dependent cells onto the desired area of an unpatterned culture substrate by micro-patterned light projection with high precision to the size of one single cell. The method is based on the phenomenon that the Ca(2+)-dependent adhesion of anchor-dependent cells becomes insensitive temporarily to the removal of Ca(2+) due to the influence of the less-invasive in situ UV irradiation. By repeating the process, a patterned coculture system can be fabricated. In clear contrast to conventional cell patterning that uses fixed-patterned substrates, the cell-retaining area is defined after cell seeding. Therefore, the cell separation on a substrate based on imaging cytometry can be implemented by combining this method with a computer-controlled microprojection coupled with a microscope. Because the cells thus patterned can continue to grow and migrate freely afterward from their initial positions, the method is expected to provide a useful cell manipulating tool, especially in cases where spontaneous development of the cells after patterning is necessary and important, such as an analysis of cell migration.
Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan.
Kimio
K
Sumaru
Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan.
Toshiyuki
T
Kanamori
D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion / Q000528:radiation effects; D006224:Cricetinae; D003412:Cricetulus; D044085:Microfluidics; D055616:Microtechnology / Q000379:methods*; D013314:Stress, Mechanical; D014466:Ultraviolet Rays
Ca(2+)-dependent adhesion; Cell migration; Cell separation; In situ micro-patterned light projection; Patterned coculture; Photoinduced enhancement of cell adhesion (PIECA)