Fabrication of gradient nanopattern by thermal nanoimprinting technique and screening of the response of human endothelial colony-forming cells

Dae Hwan Kim, Long Hui Cui, Ha Rim Seo, Hyung Joon Joo, Seung Cheol Choi, Do-Sun Lim, Kyu Back Lee

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1 Citation (Scopus)


Nanotopography can be found in various extracellular matrices (ECMs) around the body and is known to have important regulatory actions upon cellular reactions. However, it is difficult to determine the relation between the size of a nanostructure and the responses of cells owing to the lack of proper screening tools. Here, we show the development of reproducible and cost-effective gradient nanopattern plates for the manipulation of cellular responses. Using anodic aluminum oxide (AAO) as a master mold, gradient nanopattern plates with nanopillars of increasing diameter ranges [120-200 nm (GP 120/200), 200-280 nm (GP 200/280), and 280-360 nm (GP 280/360)] were fabricated by a thermal imprinting technique. These gradient nanopattern plates were designed to mimic the various sizes of nanotopography in the ECM and were used to screen the responses of human endothelial colony-forming cells (hECFCs). In this protocol, we describe the step-by-step procedure of fabricating gradient nanopattern plates for cell engineering, techniques of cultivating hECFCs from human peripheral blood, and culturing hECFCs on nanopattern plates.

Original languageEnglish
Article numbere57661
JournalJournal of Visualized Experiments
Issue number137
Publication statusPublished - 2018 Jul 1



  • Anodic aluminum oxide
  • Bioengineering
  • Extracellular matrix
  • Gradient nanopattern plates
  • Human endothelial colony-forming cells
  • Issue 137
  • Nanoimprint lithography
  • Physical stimulation

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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