A Versatile Protein and Cell Patterning Method Suitable for Long-Term Neural Cultures

Langmuir. 2019 Feb 26;35(8):2966-2975. doi: 10.1021/acs.langmuir.8b03730. Epub 2019 Feb 15.

Abstract

Herein, we present an easy-to-use protein and cell patterning method relying solely on pipetting, rinsing steps and illumination with a desktop lamp, which does not require any expensive laboratory equipment, custom-built hardware or delicate chemistry. This method is based on the adhesion promoter poly(allylamine)-grafted perfluorophenyl azide, which allows UV-induced cross-linking with proteins and the antifouling molecule poly(vinylpyrrolidone). Versatility is demonstrated by creating patterns with two different proteins and a polysaccharide directly on plastic well plates and on glass slides, and by subsequently seeding primary neurons and C2C12 myoblasts on the patterns to form islands and mini-networks. Patterning characterization is done via immunohistochemistry, Congo red staining, ellipsometry, and infrared spectroscopy. Using a pragmatic setup, patterning contrasts down to 5 μm and statistically significant long-term stability superior to the gold standard poly(l-lysine)-grafted poly(ethylene glycol) could be obtained. This simple method can be used in any laboratory or even in classrooms and its outstanding stability is especially interesting for long-term cell experiments, e.g., for bottom-up neuroscience, where well-defined microislands and microcircuits of primary neurons are studied over weeks.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biocompatible Materials / chemistry*
  • Biocompatible Materials / pharmacology*
  • Cell Culture Techniques / methods*
  • Cell Line
  • Cell Survival / drug effects
  • Myoblasts / cytology
  • Neuronal Outgrowth / drug effects
  • Neurons / cytology*
  • Neurons / drug effects*
  • Neurons / metabolism
  • Polymers / chemistry
  • Proteins / metabolism*
  • Rats
  • Surface Properties

Substances

  • Biocompatible Materials
  • Polymers
  • Proteins