Case Number 104013 - Micropatterning Two Different Cell types on Biomaterials

Contact: Geoffrey Pinski
Phone: 513-558-5696

Description:  This methodology allows direct patterning of two different cell types on biocompatible and biodegradable substrates, such as chitosan.
In this approach, an anionic cell-resistant polyelectrolyte, poly(methacry1ic acidco-oligoethyleneglycolmethacrylate) is microcontact printed as a series of 60 µm lines on a chitosan film. Monolayers of the first cell type (human vascular endothelial cells) naturally attach and proliferate only within the 20 µm lines of bare chitosan separating the 60 µm wide lines of cell resistant polyelectrolyte. The substrate is subsequently immersed into a solution of chitosan that electrostatically binds onto the 60 µm wide lines of cell-resistant polyelectrolyte, rendering these regions adhesive to a second type of cells (3T3 fibroblast cells). Optical phase contrast and fluorescence microscopy show that endothelial and fibroblast cells can be simultaneously patterned with micrometer accuracy. Cross-sectional imaging with a confocal microscope further reveals that the fibroblast cells are offset vertically by ~0.25 µm, which can be adjusted by varying the concentration of the polyelectrolyte.
This procedure is a significant advance from current micropatterning techniques that print cell-adhesive and cell-resistant patterns on non-biocompatible substrates. In comparison to other proven techniques for patterning two different cell types the polyelectrolyte assembly approach reported here is non-cytotoxic, straightforward, and allows arbitrary geometric patterns to be formed on biodegradable substrates without the need for electroactive substrates or external fields.
The organization of multiple cell types with sub-cellular resolution on biomaterials is an important first step towards the bottom-up assembly of cells to replicate tissue complexity and function that may not be possible using traditional co-culture technologies, wherein multiple cell types are randomly seeded amongst themselves. Broader and immediate applications of the approach presented here include the study of short-range surface receptor mediated heterotypic cell interactions, development of cell-based sensors, and high-throughput screening systems.