Control of Cell Adhesion and Detachment on a Nanostructured Scaffold Composed of a Light-responsive Gas Generation film

Abstract

The standard method of detaching adherent cells from a culture substrate involves the application of trypsin to digest the extracellular matrix. However, the trypsin treatment is time-consuming and sometimes causes damage to the cells when harvesting cells from a culture dish. We have developed a novel nanostructured scaffold composed of a lightresponsive gas-generating film (gas-generation nanoscaffold) that serves the dual functions of cell adhesion and cell detachment. Cell adhesion was based on the nanostructured surface topography with numerous holes of 230 nm diameter and 200 nm depth; the structure was created by soft lithography. Cell detachment was accomplished by the light irradiation of the photodecomposable polymer film to generate gas, which weakens the adhesive forces between the cells and the substrate. 11 µl of N 2 gas was generated in 90 s and the gas-generation rate was 0.1 µl/s. It was found that twice as many human bronchioalveolar carcinoma cells (NCL-H1650) adhered to the nanoscaffold than to the same scaffold without the nanostructure. The nanoscaffold exhibited no cytotoxicity according to cell viability and cytotoxicity assays. After culturing the cells on the gas-generation nanoscaffold, they were irradiated with light to generate gas, which causes the cells on the film to detach. We successfully demonstrated cell adhesion and cell detachment on the novel gas-generation nanoscaffold. This novel material is expected to be useful as an alternative cell culture substrate without the need for the standard trypsin treatment.

Contact Angle and Cell Adhesion of Micro/Nano-Structured Poly(lactic-co-glycolic acid) Membranes for Dental Regenerative Therapy

“…The growth and differentiation of C2C12 myoblasts were enhanced by grooved PLGA films (ridge/groove: 0.8 µm; depth: 0.6 µm) compared to the untreated PLGA [16]. Furthermore, two times as many human bronchioalveolar carcinoma cells adhered to a photo-decomposable polymer film with a holed structure than to the same scaffold lacking the holed structure [17]. Nano-pillars or holes have been reported to improve water repellency.…”

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

“…In a previous study, LGP films were deposited on the substrate, and cancer cells adhered to the surface were released from the vicinity of an LED (light emitting diode) light spot diameter of 6 mm. 19 However, due to the spread of bubbles and the adhesion of neighboring cells, it was difficult to release only specific cells by controlling the light. In this paper, to better control bubble generation, we developed a novel LGP microarray in which each structure was independent.…”

“…18 In previous studies, LGP has been used to release adherent cancer cells from substrates. 19 In the current study, we constructed a microarray structure using LGP, separated and arranged the cells somewhat uniformly, and then selectively retrieved target cells at a single-cell level by selective light irradiation using an inverted fluorescence microscope. The principle of the LGP microarray technique is shown in Fig.…”

Cell Culture on Low-Fluorescence and High-Resolution Photoresist

“…On the other hand, since the surface roughness affects cells and shapes of cellular tissue, cell culture on the rough surface has also received attention [32]. The surface roughness increases the strength of adhesion and support cell proliferation [33]. There is the opposite effect, depending on the cell, surface roughness reduces the adhesion area of cells and substrate, thus facilitating the formation of spheroids by promoting cell-cell adhesion [34].…”