Abstract
Lung primordial specification as well as branching morphogenesis, and the formation of various pulmonary cell lineages, requires a specific interaction of the lung endoderm with its surrounding mesenchyme and mesothelium. Lung mesenchyme has been shown to be the source of inductive signals for lung branching morphogenesis. Epithelial–mesenchymal–mesothelial interactions are also critical to embryonic lung morphogenesis. Early embryonic lung organ culture is a very useful system to study epithelial–mesenchymal interactions. Both epithelial and mesenchymal morphogenesis proceed under specific conditions that can be readily manipulated in this system (in the absence of maternal influence and blood flow). More importantly this technique can be readily done in a serumless, chemically defined culture media. Gain and loss of function can be achieved using expressed proteins, recombinant viral vectors, and/or analysis of transgenic mouse strains, antisense RNA, as well as RNA interference gene knockdown. Additionally, to further study epithelial–mesenchymal interactions, the relative roles of epithelium versus mesenchyme signaling can also be determined using tissue recombination (e.g., epithelial and mesenchymal separation) and microbead studies.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Alescio, T., and Cassini, A. (1962) Induction in vitro of tracheal buds by pulmonary mesenchyme grafted on tracheal epithelium. J. Exp. Zool. 150, 83–94.
Warburton, D., Schwarz, M., Tefft, D., Flores-Delgado, G., Anderson, K. D., and Cardoso, W. V. (2000) The molecular basis of lung morphogenesis. Mech. Dev. 92, 55–81.
Spooner, B. S., Hardman, P., and Paulsen, A. (1994) Gravity in mammalian organ development: differentiation of cultured lung and pancreas rudiments during spaceflight. J. Exp. Zool. 269, 212–222.
Jaskoll, T. F., Don-Wheeler, G., Johnson, R., and Slavkin, H. C. (1988) Embryonic mouse lung morphogenesis and type II cytodifferentiation in serumless, chemically defined medium using prolonged in vitro cultures. Cell Differ. 24, 105–117.
Seth, R., Shum, L., Wu, F., Wuenschell, C., Hall, F. L., Slavkin, H. C., and Warburton, D. (1993) Role of epidermal growth factor expression in early mouse embryo lung branching morphogenesis in culture: antisense oligodeoxynucleotide inhibitory strategy. Dev. Biol. 158, 555–559.
del Moral, P. M., De Langhe, S. P., Sala, F. G., Veltmaat, J. M., Tefft, D., Wang, K., Warburton, D., and Bellusci, S. (2006) Differential role of FGF9 on epithelium and mesenchyme in mouse embryonic lung. Dev. Biol. 293, 77–89.
Del Moral, P. M., Sala, F. G., Tefft, D., Shi, W., Keshet, E., Bellusci, S., and Warburton, D. (2006) VEGF-A signaling through Flk-1 is a critical facilitator of early embryonic lung epithelial to endothelial crosstalk and branching morphogenesis. Dev. Biol. 290, 177–188.
Deterding, R. R., and Shannon, J. M. (1995) Proliferation and differentiation of fetal rat pulmonary epithelium in the absence of mesenchyme. J. Clin. Invest. 95, 2963–2972.
Acknowledgments
This work was supported by the Saban Research Institute Pre-doctoral Award (PMDM) and by NIH RO1 HL75773.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
del Moral, PM., Warburton, D. (2010). Explant Culture of Mouse Embryonic Whole Lung, Isolated Epithelium, or Mesenchyme Under Chemically Defined Conditions as a System to Evaluate the Molecular Mechanism of Branching Morphogenesis and Cellular Differentiation. In: Ward, A., Tosh, D. (eds) Mouse Cell Culture. Methods in Molecular Biology, vol 633. Humana Press. https://doi.org/10.1007/978-1-59745-019-5_5
Download citation
DOI: https://doi.org/10.1007/978-1-59745-019-5_5
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-58829-772-3
Online ISBN: 978-1-59745-019-5
eBook Packages: Springer Protocols