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Standardization of esophageal adenocarcinoma in vitro model and its applicability for model drug testing

FLO-1 cell line represents an important tool in esophageal adenocarcinoma (EAC) research as a verified and authentic cell line to study the disease pathophysiology and antitumor drug screenings. Since in vitro characteristics of cells depend on the microenvironment and culturing conditions, we perfo...

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Autores principales: Tratnjek, Larisa, Sibinovska, Nadica, Kralj, Slavko, Makovec, Darko, Kristan, Katja, Kreft, Mateja Erdani
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7988140/
https://www.ncbi.nlm.nih.gov/pubmed/33758229
http://dx.doi.org/10.1038/s41598-021-85530-w
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author Tratnjek, Larisa
Sibinovska, Nadica
Kralj, Slavko
Makovec, Darko
Kristan, Katja
Kreft, Mateja Erdani
author_facet Tratnjek, Larisa
Sibinovska, Nadica
Kralj, Slavko
Makovec, Darko
Kristan, Katja
Kreft, Mateja Erdani
author_sort Tratnjek, Larisa
collection PubMed
description FLO-1 cell line represents an important tool in esophageal adenocarcinoma (EAC) research as a verified and authentic cell line to study the disease pathophysiology and antitumor drug screenings. Since in vitro characteristics of cells depend on the microenvironment and culturing conditions, we performed a thorough characterization of the FLO-1 cell line under different culturing conditions with the aim of (1) examining the effect of serum-free growth medium and air–liquid interface (A–L) culturing, which better reflect physiological conditions in vivo and (2) investigating the differentiation potential of FLO-1 cells to mimic the properties of the in vivo esophageal epithelium. Our study shows that the composition of the media influenced the morphological, ultrastructural and molecular characteristics of FLO-1 cells, such as the expression of junctional proteins. Importantly, FLO-1 cells formed spheres at the A–L interface, recapitulating key elements of tumors in the esophageal tube, i.e., direct contact with the gas phase and three-dimensional architecture. On the other hand, FLO-1 models exhibited high permeability to model drugs and zero permeability markers, and low transepithelial resistance, and therefore poorly mimicked normal esophageal epithelium. In conclusion, the identified effect of culture conditions on the characteristics of FLO-1 cells should be considered for standardization, data reproducibility and validity of the in vitro EAC model. Moreover, the sphere-forming ability of FLO-1 cells at the A–L interface should be considered in EAC tumor biology and anticancer drug studies as a reliable and straightforward model with the potential to increase the predictive efficiency of the current in vitro approaches.
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spelling pubmed-79881402021-03-25 Standardization of esophageal adenocarcinoma in vitro model and its applicability for model drug testing Tratnjek, Larisa Sibinovska, Nadica Kralj, Slavko Makovec, Darko Kristan, Katja Kreft, Mateja Erdani Sci Rep Article FLO-1 cell line represents an important tool in esophageal adenocarcinoma (EAC) research as a verified and authentic cell line to study the disease pathophysiology and antitumor drug screenings. Since in vitro characteristics of cells depend on the microenvironment and culturing conditions, we performed a thorough characterization of the FLO-1 cell line under different culturing conditions with the aim of (1) examining the effect of serum-free growth medium and air–liquid interface (A–L) culturing, which better reflect physiological conditions in vivo and (2) investigating the differentiation potential of FLO-1 cells to mimic the properties of the in vivo esophageal epithelium. Our study shows that the composition of the media influenced the morphological, ultrastructural and molecular characteristics of FLO-1 cells, such as the expression of junctional proteins. Importantly, FLO-1 cells formed spheres at the A–L interface, recapitulating key elements of tumors in the esophageal tube, i.e., direct contact with the gas phase and three-dimensional architecture. On the other hand, FLO-1 models exhibited high permeability to model drugs and zero permeability markers, and low transepithelial resistance, and therefore poorly mimicked normal esophageal epithelium. In conclusion, the identified effect of culture conditions on the characteristics of FLO-1 cells should be considered for standardization, data reproducibility and validity of the in vitro EAC model. Moreover, the sphere-forming ability of FLO-1 cells at the A–L interface should be considered in EAC tumor biology and anticancer drug studies as a reliable and straightforward model with the potential to increase the predictive efficiency of the current in vitro approaches. Nature Publishing Group UK 2021-03-23 /pmc/articles/PMC7988140/ /pubmed/33758229 http://dx.doi.org/10.1038/s41598-021-85530-w Text en © The Author(s) 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Tratnjek, Larisa
Sibinovska, Nadica
Kralj, Slavko
Makovec, Darko
Kristan, Katja
Kreft, Mateja Erdani
Standardization of esophageal adenocarcinoma in vitro model and its applicability for model drug testing
title Standardization of esophageal adenocarcinoma in vitro model and its applicability for model drug testing
title_full Standardization of esophageal adenocarcinoma in vitro model and its applicability for model drug testing
title_fullStr Standardization of esophageal adenocarcinoma in vitro model and its applicability for model drug testing
title_full_unstemmed Standardization of esophageal adenocarcinoma in vitro model and its applicability for model drug testing
title_short Standardization of esophageal adenocarcinoma in vitro model and its applicability for model drug testing
title_sort standardization of esophageal adenocarcinoma in vitro model and its applicability for model drug testing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7988140/
https://www.ncbi.nlm.nih.gov/pubmed/33758229
http://dx.doi.org/10.1038/s41598-021-85530-w
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