Microscale Control of Cell Contact and Spacing via Three-Component Surface Patterning

Abstract

The complexity of micropatterned cell constructs has been limited by difficulties in patterning more than two surface components on a culture substrate. Photolithography using multiple aligned masks is well established for generalized multicomponent patterning, but is often too harsh for biomolecules. We report a two-mask photolithographic process that is tuned to preserve bioactivity in patterns composed of covalently coupled polyethylene glycol (PEG), adsorbed extracellular matrix protein (e.g. collagen I), and adsorbed serum proteins (e.g. vitronectin). Thereby, we pattern two cell types-primary hepatocytes and 3T3 fibroblasts-demonstrating control over contact and spacing (20-200 μm) between the two cell types for over one week. This method is applicable to the study of intercellular communication in cell biology and tissue engineering.

Targeting liver fibrosis: Strategies for development and validation of antifibrotic therapies

“…Thus, a controlled 3D coculture system using hepatocytes and 3T3 fibroblasts was recently reported. 33 Projects to create multicellular artificial cocultures of hepatocytes, HSC, endothelial cells, Kupffer cells, and cholangiocytes are under way and may provide a highly reproducible and cost-efficient platform to develop and test antifibrotic drugs in a nearly physiological hepatic microenvironment.…”

Microfabrication of Patterned Co-cultures for Controllable Cell–Cell Interfaces

“…In the case of negative photoresists, only the exposed regions remain after developing while the rest is washed off. For instance, micropatterned co-cultures of hepatocyte and 3T3-J2 cells were fabricated by patterning collagen and poly (ethylene glycol) (PEG), an antifouling polymer, on the culture substrate [ 14 ]. In this study, collagen was adsorbed and then patterned on the surface by the photoresist lift-off process.…”

Characterization of microfluidic human epidermal keratinocyte culture

“…The use of HEK in toxicity assays (Chou et al 2002;Didier et al 1999;Monteiro-Riviere et al 2005;Rouse et al 2006;Shvedova et al 2003) makes this cell type an attractive candidate for use in microfluidic high throughput toxicity assays. Cell culture in microfluidic systems has been demonstrated by others Delamarche et al 2005;Figallo et al 2007;Flaim et al 2005;Folch and Toner 2000;Gu et al 2004;Hui and Bhatia 2007;Hung et al 2005a, Hung et al b;Kane et al 2006;Khademhosseini et al 2004;Kim et al 2006;Leclerc et al 2006a;b;Lee et al 2006;Prokop et al 2004;Raty et al 2004;Rhee et al 2005;Sin et al 2004;Song et al 2005;Toh et al 2007;Tourovskaia et al 2005;Walker et al 2002) but keratinocytes have not yet been studied at the microscale.…”