Simple benchtop patterning of hydrogel grids for living cell microarrays

Abstract

A living cell microarray consists of an orderly arrangement of cells attached to a solid support such as a glass microscope slide. The chief difficulty of obtaining such arrays is the fabrication of substrates patterned with micro-wells, adhesive spots, or other features to guide orderly cell attachment. Here we report a novel method using woven Nylon mesh to micropattern three-dimensional alginate hydrogel grids on glass slides. The Nylon mesh is both inexpensive and off-the-shelf. By using Nylon mesh we have eliminated any need for lithography, clean room equipment, and microarray printers to generate microarray patterns; thus, this technique can be easily adopted by biological research labs that may lack microfabrication expertise and facilities. We have demonstrated that glass slides micropatterned with hydrogel grids guide the orderly attachment of single fibroblast cells and Schwann cell clusters in microarrays. The fibroblast arrays consisted of 70 µm square compartments at a density of 21 000 compartments per cm². The Schwann cell arrays consisted of 100 µm square compartments at a density of 6000 per cm². This patterning technique addresses the need for a simple, inexpensive, benchtop method for micro-patterning glass slides and obtaining living cell microarrays.