Mammalian cell division in 3D matrices via quantitative confocal reflection microscopy

Lijuan He, Alexandra Sneider, Weitong Chen, Michelle Karl, Vishnu Prasath, Pei Hsun Wu, Gunnar Mattson, Denis Wirtz

Research output: Contribution to journalArticle

Abstract

The study of how mammalian cell division is regulated in a 3D environment remains largely unexplored despite its physiological relevance and therapeutic significance. Possible reasons for the lack of exploration are the experimental limitations and technical challenges that render the study of cell division in 3D culture inefficient. Here, we describe an imaging-based method to efficiently study mammalian cell division and cellmatrix interactions in 3D collagen matrices. Cells labeled with fluorescent H2B are synchronized using the combination of thymidine blocking and nocodazole treatment, followed by a mechanical shake-off technique. Synchronized cells are then embedded into a 3D collagen matrix. Cell division is monitored using live-cell microscopy. The deformation of collagen fibers during and after cell division, which is an indicator of cell-matrix interaction, can be monitored and quantified using quantitative confocal reflection microscopy. The method provides an efficient and general approach to study mammalian cell division and cell-matrix interactions in a physiologically relevant 3D environment. This approach not only provides novel insights into the molecular basis of the development of normal tissue and diseases, but also allows for the design of novel diagnostic and therapeutic approaches.

Original languageEnglish (US)
Article numbere56364
JournalJournal of Visualized Experiments
Volume2017
Issue number129
DOIs
StatePublished - Nov 29 2017

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Keywords

  • 3D matrix
  • Bioengineering
  • Cell division
  • Cell-matrix interaction
  • Confocal reflection microscopy
  • Issue 129
  • Synchronization

ASJC Scopus subject areas

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

Cite this

He, L., Sneider, A., Chen, W., Karl, M., Prasath, V., Wu, P. H., Mattson, G., & Wirtz, D. (2017). Mammalian cell division in 3D matrices via quantitative confocal reflection microscopy. Journal of Visualized Experiments, 2017(129), [e56364]. https://doi.org/10.3791/56364