Aqueous Processed Biopolymer Interfaces for Single-Cell Microarrays

Vittorio Ferrara,Valeria Vetri,Giuseppe Arrabito,Carla Giordano,Sebastiano Cataldo,Michelangelo Scopelliti,Giovanni Zito,Bruno Pignataro

doi:10.1021/acsbiomaterials.9b01871

Vittorio Ferrara, Valeria Vetri + Show 6 more

Open Access

https://doi.org/10.1021/acsbiomaterials.9b01871

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Abstract

Single-cell microarrays are emerging tools to unravel intrinsic diversity within complex cell populations, opening up new approaches for the in-depth understanding of highly relevant diseases. However, most of the current methods for their fabrication are based on cumbersome patterning approaches, employing organic solvents and/or expensive materials. Here, we demonstrate an unprecedented green-chemistry strategy to produce single-cell capture biochips onto glass surfaces by all-aqueous inkjet printing. At first, a chitosan film is easily inkjet printed and immobilized onto hydroxyl-rich glass surfaces by electrostatic immobilization. In turn, poly(ethylene glycol) diglycidyl ether is grafted on the chitosan film to expose reactive epoxy groups and induce antifouling properties. Subsequently, microscale collagen spots are printed onto the above surface to define the attachment area for single adherent human cancer cells harvesting with high yield. The reported inkjet printing approach enables one to modulate the collagen area available for cell attachment in order to control the number of captured cells per spot, from single-cells up to double- and multiple-cell arrays. Proof-of-principle of the approach includes pharmacological treatment of single-cells by the model drug doxorubicin. The herein presented strategy for single-cell array fabrication can constitute a first step toward an innovative and environmentally friendly generation of aqueous-based inkjet-printed cellular devices.

Highlights

The investigation of cellular systems at the single-cell level, i.e., single-cell biology, permits one to shed light on their relevant biochemical and biophysical processes at an unprecedented level of detail.[1,2] Typically, populations of cells are investigated in standard cell cultures conditions, so the resulting extracted biological information does often consist of a broad average output from a cell population
Single-cell investigation approaches provide the ultimate level of resolution in our quest to capture relevant heterogeneities, constituting versatile tools for disease biomarkers identification,[3] drug discovery,[5] intracellular fluorescence-based molecular tracking,[6] and stochastic gene expression.[7]
Single cells have been caught by microfluidic approaches onto functionalized materials, whose surfaces have been chemically tailored, in order to introduce cell-adhesive properties and biological selectivity.[14,15]

Summary

INTRODUCTION

The investigation of cellular systems at the single-cell level, i.e., single-cell biology, permits one to shed light on their relevant biochemical and biophysical processes at an unprecedented level of detail.[1,2] Typically, populations of cells are investigated in standard cell cultures conditions, so the resulting extracted biological information does often consist of a broad average output from a cell population. Nonadherent single-cells microarrays were obtained by physically trapping individual cells in polymeric microchambers that can accommodate only one individual cell per well.[12,13] single cells have been caught by microfluidic approaches onto functionalized materials, whose surfaces have been chemically tailored, in order to introduce cell-adhesive properties and biological selectivity.[14,15] An even further control of the spatial arrangement and material composition of the cell-capture features becomes crucial allowing for the development of functional single-cell biology platforms In this regard, printing techniques allow for the direct fabrication of a large variety of biomolecular structures at micro- or nanoscale resolution and have, been extensively pursued to obtain adherent single-cells microarrays.[16,17] For instance, microcontact printing and scanning probe-based methods, such as dip pen nanolithography and polymer pen lithography, allow for the direct fabrication of cell-adhesion micropatterns to control the attachment area, inducing stimuli on single cells down to the subcellular scale (

EXPERIMENTAL SECTION

RESULTS AND DISCUSSION

CONCLUSIONS

■ ACKNOWLEDGMENTS

■ REFERENCES