Culturing MDCK cells in three dimensions for analyzing intracellular dynamics.
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.
Elia
Natalie
N
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland.
Lippincott-Schwartz
Jennifer
J
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland.
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D016764:Cell Polarity; D003593:Cytoplasm / Q000648:ultrastructure; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology* / Q000276:immunology; D036681:Fluorescence Recovery After Photobleaching; D019084:Fluorescent Antibody Technique, Indirect; D021621:Imaging, Three-Dimensional; D007668:Kidney / Q000166:cytology* / Q000276:immunology; D018613:Microscopy, Confocal; D009024:Morphogenesis; D023822:Tissue Engineering