UNIT 4.22 Culturing MDCK Cells in Three Dimensions for Analyzing Intracellular Dynamics

  1. Natalie Elia,
  2. Jennifer Lippincott-Schwartz

Published Online: 1 JUN 2009

DOI: 10.1002/0471143030.cb0422s43

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Elia, N. and Lippincott-Schwartz, J. 2009. Culturing MDCK Cells in Three Dimensions for Analyzing Intracellular Dynamics. Current Protocols in Cell Biology. 43:4.22:4.22.1–4.22.18.

Author Information

  1. Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland

Publication History

  1. Published Online: 1 JUN 2009
  2. Published Print: JUN 2009


Epithelial cells grown in three-dimensional (3-D) cultures of extracellular matrix differentiate into a multicellular structure of polarized cells. This process shares many characteristics with the physiological development of an epithelial tissue and the formation of polarity in epithelial cells. Imaging 3-D cultures of polarized epithelial cells is therefore a powerful tool to study epithelial architecture and morphogenesis under close-to-physiological conditions. The new generation of confocal microscopes allows live-cell imaging of fluorescently tagged molecules in these cultures. This opens up new opportunities for studying how molecules behave and are distinguished asymmetrically within a 3-D setting. This unit discusses technical aspects for culturing and imaging MDCK 3-D culture for both fixed 3-D cultures and live-cell imaging. Curr. Protoc. Cell Biol. 43:4.22.1-4.22.18. © 2009 by John Wiley & Sons, Inc.


  • 3-D cultures;
  • MDCK cells;
  • live-cell imaging;
  • MDCK cyst;
  • epithelial cyst;
  • 3-D culture imaging