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@inbook{1394952,
author = {Rabata, Anas and Hampl, Aleš and Koledová, Zuzana},
address = {New York},
booktitle = {3D Cell Culture : Methods and Protocols},
doi = {http://dx.doi.org/10.1007/978-1-4939-7021-6_11},
edition = {1st. ed.},
editor = {Koledova Zuzana},
keywords = {3D culture; Lung epithelial stem/progenitor cells; Lungosphere assay; Spheroid culture},
howpublished = {tištěná verze "print"},
language = {eng},
location = {New York},
isbn = {978-1-4939-7019-3},
pages = {149-165},
publisher = {Humana Press},
title = {Lungosphere Assay: 3D Culture of Lung Epithelial Stem/Progenitor Cells},
url = {https://link.springer.com/protocol/10.1007/978-1-4939-7021-6_11},
year = {2017}
}
TY - CHAP
ID - 1394952
AU - Rabata, Anas - Hampl, Aleš - Koledová, Zuzana
PY - 2017
TI - Lungosphere Assay: 3D Culture of Lung Epithelial Stem/Progenitor Cells
VL - Methods in Molecular Biology, volume 1612
PB - Humana Press
CY - New York
SN - 9781493970193
KW - 3D culture
KW - Lung epithelial stem/progenitor cells
KW - Lungosphere assay
KW - Spheroid culture
UR - https://link.springer.com/protocol/10.1007/978-1-4939-7021-6_11
L2 - https://link.springer.com/protocol/10.1007/978-1-4939-7021-6_11
N2 - Lung epithelium contains distinctive subpopulations of lung stem/progenitor cells (LSPCs) that are essential for lung epithelial maintenance and repair in vivo. Hence, LSPCs are in the center of interest of lung biology due to their promising therapeutic applications. To reach this goal, proper characterization of LSPCs, understanding of their proliferation and differentiation potentials and elucidation of mechanisms that control them are necessary. Therefore, development of reliable in vitro clonogenic assays has been needed. We established lungosphere assay, an in vitro sphere-forming 3D culture assay that enables to evaluate stem/progenitor cell activity, self-renewal and differentiation capacity of LSPCs and to conveniently test the effect of various treatments on LSPCs. Here we provide a detailed description of procedures for isolation of adult mouse lung epithelial cells, their culture in non-adherent conditions to form LSPC-derived spheroids (lungospheres) and for embedding of lungospheres into 3D extracellular matrix to model processes of lung tissue maintenance in a physiologically relevant microenvironment.
ER -
RABATA, Anas, Aleš HAMPL and Zuzana KOLEDOVÁ. Lungosphere Assay: 3D Culture of Lung Epithelial Stem/Progenitor Cells. In Koledova Zuzana. \textit{3D Cell Culture : Methods and Protocols}. 1st. ed. New York: Humana Press. p.~149-165. Methods in Molecular Biology, volume 1612. ISBN~978-1-4939-7019-3. doi:10.1007/978-1-4939-7021-6\_{}11. 2017.
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