Aqueous Processed Biopolymer Interfaces for Single-Cell Microarrays

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.

An electroactive hybrid biointerface for enhancing neuronal differentiation and axonal outgrowth on bio-subretinal chip

“…Given that the needle size employed in this study was the smallest available size by the manufacturer, future studies can divert towards non-contact droplets printing methods such as piezoelectric inkjet printing or drop-impact printing systems, which could significantly reduce the minimum limit of micropattern size [ 59 , 60 ]. Several advanced bioprinting systems have recently demonstrated the possibility of producing cell micropatterns within 30 ​μm using drop-on-demand inkjet printing [ 61 , 62 ]. Notably, we also attempted to directly print cells by incorporating the PC-12 ​cells into the aGO-COL composite at a concentration of 1 ​× ​10 7 ​cells/mL as a bioink ( Fig.…”

2D and 3D inkjet printing of biopharmaceuticals – A review of trends and future perspectives in research and manufacturing

“…Instead of direct dispensing of cells into a specific position, a surface is modified digitally to provide sites for cell attachment. This method removes the need to expose cells to the potentially damaging effects of inkjet printing [50], [51] but relies on the reliability of single cell attachment, with examples showing yields from 60% [52] to 80% [53].…”

Artificial Biosystems by Printing Biology

“…[118,119] 2D inkjet-printed chitosan patterns have also been employed as structural layer for the fabrication of single-cell arrays. [120] 3D printed chitosan scaffolds have also been loaded with bioactive molecules, to develop an artificial printed architecture for cell adhesion scaffold and surface-mediated drug delivery system. [121] Also, negatively charged polysaccharides have been exploited as structural matrix for artificial tissue constructs, as reported for gels of alginate.…”