Recent Advances in Cell Micropatterning Techniques for Bioanalytical and Biomedical Sciences

Abstract

Cell micropatterning is a method for controlling the placement of living cells on a substrate surface. [1][2][3][4][5][6][7][8] It is important for a wide range of applications, such as tissue engineering, cellbased drug screening, and fundamental cell biology studies. Most cell micropatterning methods fall into three categories based on strategy: (1) seeding cells on a chemically patterned surface of different cell adhesiveness, (2) seeding cells on a topographically patterned surface, or (3) directed delivery of cells onto discrete regions of a substrate. Although the latter two categories are also important, this review mainly focuses on the first category. This is because it is the most common strategy and it provides tools to study fundamental aspects of cell adhesion at the single protein/receptor molecule level. 9There are excellent reviews that also cover the latter two categories. 3,8,10 We first introduce some of the important applications of cell micropatterning in general. We then describe methods for micropatterning the cell adhesiveness, referring to materials that promote or inhibit cell adhesion. We then move on to much more sophisticated substrates, so-called "dynamic substrates", whose cell adhesiveness can be changed by an external stimulus, such as heat, voltage and light. As an example of dynamic substrates, we describe a caged culture substrate developed by our group. This type of functional substrate opens up new possibilities in bioanalytical and biomedical sciences. Cell micropatterning is an important technique for a wide range of applications, such as tissue engineering, cell-based drug screening, and fundamental cell biology studies. This paper overviews cell patterning techniques based on chemically modified substrates with different degrees of cell adhesiveness. In particular, the focus is on dynamic substrates that change their cell adhesiveness in response to external stimuli, such as heat, voltage, and light. Such substrates allow researchers to achieve an in situ alteration of patterns of cell adhesiveness, which is useful for coculturing multiple cell types and analyzing dynamic cellular activities. As an example of dynamic substrates, we introduce a dynamic substrate based on a caged compound, where we accomplished a light-driven alteration of cell adhesiveness and the analysis of a single cell's motility.

Reprogramming cell shape with laser nano-patterning

“…Several approaches have been used to overcome this limitation and alter the adhesive environment surrounding the micropatterns on which living cells are attached (Nakanishi et al, 2008). Electric potential has been used to detach cell-repellent coatings, either by detaching micropatterned electroactive groups (Raghavan et al, 2010) or by desorbing coatings on electrodes (Gabi et al, 2010;Kaji et al, 2006), thereby allowing constrained multicellular groups of cells on large micropatterns to specifically invade the activated regions.…”

Dynamic culture substrate that captures a specific extracellular matrix protein in response to light

“…Moreover, functionalized substrates that capture or expose a cell-adhesive peptide on their surfaces in response to an external stimulus, such as heat [5], voltage [6][7][8] or light [9][10][11], are useful for the dynamic control of cell adhesion. These substrates, called dynamic substrates, have been applied not only to explore cell migration but also to coculture heterotypic cells [12]. Almost all of the earlier dynamic substrates used linear or cyclic RGD peptides as cell-adhesive ligands.…”

Micropatterning as a tool to decipher cell morphogenesis and functions

“…To that end, technological efforts have been made to fabricate so-called dynamic or switchable surfaces, whose adhesiveness can be turned on and off at will (see panel A). Several stimuli can be used to regulate the physico-chemical properties of a substrate in a localised manner, including electric potential, temperature, pH or light (reviewed in Liu et al, 2005a;Nakanishi et al, 2008). Electric fields can be used to detach the cell-repellent surfaces (Fan et al, 2008;Yeo and Mrksich, 2006;Yousaf et al, 2001).…”