Covalent Microcontact Printing of Proteins for Cell Patterning

Abstract

We describe a straightforward approach to the covalent immobilization of cytophilic proteins by microcontact printing, which can be used to pattern cells on substrates. Cytophilic proteins are printed in micropatterns on reactive self-assembled monolayers by using imine chemistry. An aldehyde-terminated monolayer on glass or on gold was obtained by the reaction between an amino-terminated monolayer and terephthaldialdehyde. The aldehyde monolayer was employed as a substrate for the direct microcontact printing of bioengineered, collagen-like proteins by using an oxidized poly(dimethylsiloxane) (PDMS) stamp. After immobilization of the proteins into adhesive "islands", the remaining areas were blocked with amino-poly(ethylene glycol), which forms a layer that is resistant to cell adhesion. Human malignant carcinoma (HeLa) cells were seeded and incubated onto the patterned substrate. It was found that these cells adhere to and spread selectively on the protein islands, and avoid the poly(ethylene glycol) (PEG) zones. These findings illustrate the importance of microcontact printing as a method for positioning proteins at surfaces and demonstrate the scope of controlled surface chemistry to direct cell adhesion.

A generic expression system to produce proteins that co-assemble with alkane thiol SAM

“…The ability of phospholipid layers to limit nonspecifi c binding has been exploited in combination with patterned arrays of soluble proteins (Kung et al 2000), whilst other approaches have combined the peptide with the lipid itself (Svedhem et al 2003) or printed proteins onto a reactive SAM (Rozkiewicz et al 2006). Incorporation of functional synthetic peptides into SAM is straightforward and patterning has been achieved by micro-contact printing ethylene glycol-terminated SAM in patterns leaving exposed gold for subsequent peptide assembly (Zhang et al 1999).…”

Extracellular-controlled breast cancer cell formation and growth using non-UV patterned hydrogels via optically-induced electrokinetics

“…The most often used traditional method for cell patterning is based on microcontact printing. [8][9][10] In this method, extracellular matrix proteins, such as actin, fibronectin, and collagen are grafted onto a biocompatible, but bio-passivated, substrate in a pre-defined pattern utilizing a master mold, such as a polydimethylsiloxane (PDMS) stamp with corresponding micron-scale features. Due to the bio-adhesive nature of these proteins, cells would be attracted to, and grow on, these areas.…”

Nanofabrication of bioinspired architectures with light harvesting proteins

“…73 Composite stamps produced from two UV-curable materials, 72 and poly(ethylene glycol) diacrylate, were used to perform μCP of polar biomolecules. 78 μCP has been used to direct the adsorption of cells on surfaces. [75][76][77] For example HeLa cells were patterned using the interaction with patterned Col3a1 protein.…”

“…[75][76][77] For example HeLa cells were patterned using the interaction with patterned Col3a1 protein. 78 Silicon oxide substrates were modified with amino-terminated SAMs and then the amino groups were converted into aldehyde groups by the reaction with terephthaldialdehyde. 79 Substrates modified in this manner can be directly patterned with collagen-like proteins by microcontact printing using a PDMS stamp ( Figure 1.4a).…”