Fabricating complex culture substrates using robotic microcontact printing (R-µCP) and sequential nucleophilic substitution.

Autor: Knight GT; Department of Biomedical Engineering, University of Wisconsin, Madison., Klann T; Department of Biomedical Engineering, University of Wisconsin, Madison., McNulty JD; Department of Biomedical Engineering, University of Wisconsin, Madison; Department of Mechanical Engineering, University of Wisconsin, Madison., Ashton RS; Department of Biomedical Engineering, University of Wisconsin, Madison; rashton2@wisc.edu.
Jazyk: angličtina
Zdroj: Journal of visualized experiments : JoVE [J Vis Exp] 2014 Oct 31 (92), pp. e52186. Date of Electronic Publication: 2014 Oct 31.
DOI: 10.3791/52186
Abstrakt: In tissue engineering, it is desirable to exhibit spatial control of tissue morphology and cell fate in culture on the micron scale. Culture substrates presenting grafted poly(ethylene glycol) (PEG) brushes can be used to achieve this task by creating microscale, non-fouling and cell adhesion resistant regions as well as regions where cells participate in biospecific interactions with covalently tethered ligands. To engineer complex tissues using such substrates, it will be necessary to sequentially pattern multiple PEG brushes functionalized to confer differential bioactivities and aligned in microscale orientations that mimic in vivo niches. Microcontact printing (μCP) is a versatile technique to pattern such grafted PEG brushes, but manual μCP cannot be performed with microscale precision. Thus, we combined advanced robotics with soft-lithography techniques and emerging surface chemistry reactions to develop a robotic microcontact printing (R-μCP)-assisted method for fabricating culture substrates with complex, microscale, and highly ordered patterns of PEG brushes presenting orthogonal 'click' chemistries. Here, we describe in detail the workflow to manufacture such substrates.
Databáze: MEDLINE