Selective cell retrieval method using light-responsive gas-generating polymer-based microarrays

Abstract

Selective cell retrieval from base material is necessary for developing and improving cell analyzing technologies as well as regenerative medicine. Many conventional technologies, such as micromanipulators, are developed for selective cell retrieval. However, selective cell retrieval at the single-cell level remains challenging because it is quite difficult to retrieve adhered single cells from base material with ease, rapidity, and no damage. Here, we propose a novel selective cell retrieval method using microarrays made of a light-responsive gas-generating polymer (LGP microarray). The convex LGP microarray was fabricated by a molding process using polystyrene microarray chips. LGP microarrays generate N₂ gas when exposed to a specific light used for fluorescence microscopy. A human cervical cancer cell (HeLa) suspension was spread on the LGP microarray coated by fibronectin. After these HeLa cells were adhered to the surface of the LGP microarray structure, light at a wavelength of 365 nm was used to irradiate the LGP microarray. All the target HeLa cells were selectively released from the light-irradiated surface area of the LGP microarray by the generated N₂ gas. The LGP microarray system was also applied to single-cell retrieval, and we easily and rapidly retrieved 100% of the single HeLa cells from the microarrays. In addition, approximately 90% of single HeLa cells retrieved from the LGP microarray proliferated on a chamber of a 96-well plate. Therefore, the LGP microarray system enables easy and selective retrieval of adhered cell groups or single cells with only harmless light irradiation.