Statistical analysis of immuno-functionalized tumor-cell behaviors on nanopatterned substrates

Dong-Joo Kim,Sang-Kwon Lee,Gil-Sung Kim,Geehee Lee

doi:10.1186/1556-276x-7-637

Dong-Joo Kim, Sang-Kwon Lee + Show 2 more

Open Access

https://doi.org/10.1186/1556-276x-7-637

Copy DOI

Abstract

Laser scanning cytometry has been proven as a powerful technology for high-content, high-throughput quantitative analysis of cellular functions in a fully automated manner. It utilizes a large-area fluorescence imaging scheme and rigorous image quantitation algorithms to enable informative analysis of cell samples attached to solid substrates. While this technology represents a powerful approach for high-content screening using cell lines, it has not been applied to the study of tumor-cell behaviors on these solid nanopatterned substrates after several hours of incubation. Herein, we statistically demonstrated functional cellular morphology information, including size, shape, and distribution of the captured cells after 0.5 to 45 h of incubation on nanopatterned substrates, such as silicon nanowires and quartz nanopillars, along with planar glass substrates. With increasing incubation time up to 45 h, we observed that the nanopatterned substrates could have not only increased adhesion and traction forces between cells and nanopatterned substrates, but also limited cell spreading on the substrates compared to the planar glass substrates. On the basis of our results, we suggest that the most important factors to influence the cell behaviors on the three solid substrates are the degree of dimension on cell behaviors and cell traction force.

Highlights

Nanostructures have been increasingly used for studies on cell interaction with solid nanostructures because unique properties of nanostructured surfaces enable a variety of novel functions of immobilized cells on the nanostructured surfaces [1,2,3]
We have demonstrated a novel platform for separating CD4+ T lymphocytes from mouse splenocytes using streptavidin (STR)-functionalized and vapor–liquid-solid-grown silicon nanowire (SiNW) [5] as well as transparent quartz nanopillar (QNP) arrays [8] having a higher separation
CA, USA) and reacted at 4°C for 20 min on STR-functionalized substrates (SiNW, QNP, and planar glass) prior to loading the A549 cells, which were manually counted using a hemocytometer with approximately 10% error (Hausser Scientific Co., PA, USA), into the PDMS cell-capture chambers (Figure 1c)

Summary

Introduction

Nanostructures have been increasingly used for studies on cell interaction with solid nanostructures because unique properties of nanostructured surfaces enable a variety of novel functions of immobilized cells on the nanostructured surfaces [1,2,3]. Owing to high surface area and increased nanowire-cell surface interaction, nanowire arrays functionalized with capture agents were demonstrated for high-yield capture of surface-bound cells including immune cell subsets and circulating tumor cells (CTCs) [5,6,7]. We have demonstrated a novel platform for separating CD4+ T lymphocytes from mouse splenocytes using streptavidin (STR)-functionalized and vapor–liquid-solid-grown silicon nanowire (SiNW) [5] as well as transparent quartz nanopillar (QNP) arrays [8] having a higher separation.

Methods

Results

Conclusion