Generation of a Close-to-Native In Vitro System to Study Lung
Cells-Extracellular Matrix Crosstalk

J 2018

Generation of a Close-to-Native In Vitro System to Study Lung Cells-Extracellular Matrix Crosstalk

GARLÍKOVÁ, Zuzana, Ana Catarina SILVA, Anas RABATA, David POTĚŠIL, Ivana IHNATOVÁ et. al.

Basic information

Original name

Generation of a Close-to-Native In Vitro System to Study Lung Cells-Extracellular Matrix Crosstalk

Authors

GARLÍKOVÁ, Zuzana (203 Czech Republic, belonging to the institution), Ana Catarina SILVA (620 Portugal), Anas RABATA (760 Syrian Arab Republic, belonging to the institution), David POTĚŠIL (203 Czech Republic, belonging to the institution), Ivana IHNATOVÁ (703 Slovakia, belonging to the institution), Jana DUMKOVÁ (203 Czech Republic, belonging to the institution), Zuzana KOLEDOVÁ (703 Slovakia, belonging to the institution), Zbyněk ZDRÁHAL (203 Czech Republic, belonging to the institution), Vladimír VINARSKÝ (203 Czech Republic), Aleš HAMPL (203 Czech Republic, guarantor, belonging to the institution), Perpétua PINTO-DO-Ó (620 Portugal) and Diana Santos NASCIMENTO (620 Portugal)

Edition

TISSUE ENGINEERING PART C-METHODS, New Rochelle, Mary Ann Liebert, 2018, 1937-3384

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10601 Cell biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 2.638

RIV identification code

RIV/00216224:14110/18:00100743

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.1089/ten.tec.2017.0283

UT WoS

000414350600001

Keywords in English

decellularization; lung; extracellular matrix; in vitro models; lung fibroblasts; biological scaffold

Abstract

V originále

Extracellular matrix (ECM) is an essential component of the tissue microenvironment, actively shaping cellular behavior. In vitro culture systems are often poor in ECM constituents, thus not allowing for naturally occurring cell–ECM interactions. This study reports on a straightforward and efficient method for the generation of ECM scaffolds from lung tissue and its subsequent in vitro application using primary lung cells. Mouse lung tissue was subjected to decellularization with 0.2% sodium dodecyl sulfate, hypotonic solutions, and DNase. Resultant ECM scaffolds were devoid of cells and DNA, whereas lung ECM architecture of alveolar region and blood and airway networks were preserved. Scaffolds were predominantly composed of core ECM and ECM-associated proteins such as collagens I-IV, nephronectin, heparan sulfate proteoglycan core protein, and lysyl oxidase homolog 1, among others. When homogenized and applied as coating substrate, ECM supported the attachment of lung fibroblasts (LFs) in a dose-dependent manner. After ECM characterization and biocompatibility tests, a novel in vitro platform for three-dimensional (3D) matrix repopulation that permits live imaging of cell–ECM interactions was established. Using this system, LFs colonized the ECM scaffolds, displaying a close-to-native morphology in intimate interaction with the ECM fibers, and showed nuclear translocation of the mechanosensor yes-associated protein (YAP), when compared with cells cultured in two dimensions. In conclusion, we developed a 3D-like culture system, by combining an efficient decellularization method with a live-imaging culture platform, to replicate in vitro native lung cell–ECM crosstalk. This is a valuable system that can be easily applied to other organs for ECM-related drug screening, disease modeling, and basic mechanistic studies.

Links

GBP206/12/G151, research and development project

Name: Centrum nových přístupů k bioanalýze a molekulární diagnostice

LM2015043, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

LM2015062, research and development project

Name: Národní infrastruktura pro biologické a medicínské zobrazování

Investor: Ministry of Education, Youth and Sports of the CR

MUNI/A/1369/2016, interní kód MU

Name: Zdroje pro tkáňové inženýrství 7 (Acronym: TissueENG 7)

Investor: Masaryk University, Category A

NV16-31501A, research and development project

Name: Tkáňové inženýrství epitelů: Buňky a protokoly pro regenerativní medicínu

Citovat

GARLÍKOVÁ, Zuzana, Ana Catarina SILVA, Anas RABATA, David POTĚŠIL, Ivana IHNATOVÁ, Jana DUMKOVÁ, Zuzana KOLEDOVÁ, Zbyněk ZDRÁHAL, Vladimír VINARSKÝ, Aleš HAMPL, Perpétua PINTO-DO-Ó and Diana Santos NASCIMENTO. Generation of a Close-to-Native In Vitro System to Study Lung Cells-Extracellular Matrix Crosstalk. TISSUE ENGINEERING PART C-METHODS. New Rochelle: Mary Ann Liebert, 2018, vol. 24, No 1, p. 1-13. ISSN 1937-3384. Available from: https://dx.doi.org/10.1089/ten.tec.2017.0283.

@article{1394260,
   author = {Garlíková, Zuzana and Silva, Ana Catarina and Rabata, Anas and Potěšil, David and Ihnatová, Ivana and Dumková, Jana and Koledová, Zuzana and Zdráhal, Zbyněk and Vinarský, Vladimír and Hampl, Aleš and PintoanddoandÓ, Perpétua and Nascimento, Diana Santos},
   article_location = {New Rochelle},
   article_number = {1},
   doi = {http://dx.doi.org/10.1089/ten.tec.2017.0283},
   keywords = {decellularization; lung; extracellular matrix; in vitro models; lung fibroblasts; biological scaffold},
   language = {eng},
   issn = {1937-3384},
   journal = {TISSUE ENGINEERING PART C-METHODS},
   title = {Generation of a Close-to-Native In Vitro System to Study Lung Cells-Extracellular Matrix Crosstalk},
   url = {http://online.liebertpub.com/doi/abs/10.1089/ten.tec.2017.0283?url_ver=Z39.88-2003&rfr_id=ori%3Arid%3Acrossref.org&rfr_dat=cr_pub%3Dpubmed},
   volume = {24},
   year = {2018}
}

TY  - JOUR
ID  - 1394260
AU  - Garlíková, Zuzana - Silva, Ana Catarina - Rabata, Anas - Potěšil, David - Ihnatová, Ivana - Dumková, Jana - Koledová, Zuzana - Zdráhal, Zbyněk - Vinarský, Vladimír - Hampl, Aleš - Pinto-do-Ó, Perpétua - Nascimento, Diana Santos
PY  - 2018
TI  - Generation of a Close-to-Native In Vitro System to Study Lung Cells-Extracellular Matrix Crosstalk
JF  - TISSUE ENGINEERING PART C-METHODS
VL  - 24
IS  - 1
SP  - 1-13
EP  - 1-13
PB  - Mary Ann Liebert
SN  - 19373384
KW  - decellularization
KW  - lung
KW  - extracellular matrix
KW  - in vitro models
KW  - lung fibroblasts
KW  - biological scaffold
UR  - http://online.liebertpub.com/doi/abs/10.1089/ten.tec.2017.0283?url_ver=Z39.88-2003&rfr_id=ori%3Arid%3Acrossref.org&rfr_dat=cr_pub%3Dpubmed
N2  - Extracellular matrix (ECM) is an essential component of the tissue microenvironment, actively shaping cellular behavior. In vitro culture systems are often poor in ECM constituents, thus not allowing for naturally occurring cell–ECM interactions. This study reports on a straightforward and efficient method for the generation of ECM scaffolds from lung tissue and its subsequent in vitro application using primary lung cells. Mouse lung tissue was subjected to decellularization with 0.2% sodium dodecyl sulfate, hypotonic solutions, and DNase. Resultant ECM scaffolds were devoid of cells and DNA, whereas lung ECM architecture of alveolar region and blood and airway networks were preserved. Scaffolds were predominantly composed of core ECM and ECM-associated proteins such as collagens I-IV, nephronectin, heparan sulfate proteoglycan core protein, and lysyl oxidase homolog 1, among others. When homogenized and applied as coating substrate, ECM supported the attachment of lung fibroblasts (LFs) in a dose-dependent manner. After ECM characterization and biocompatibility tests, a novel in vitro platform for three-dimensional (3D) matrix repopulation that permits live imaging of cell–ECM interactions was established. Using this system, LFs colonized the ECM scaffolds, displaying a close-to-native morphology in intimate interaction with the ECM fibers, and showed nuclear translocation of the mechanosensor yes-associated protein (YAP), when compared with cells cultured in two dimensions. In conclusion, we developed a 3D-like culture system, by combining an efficient decellularization method with a live-imaging culture platform, to replicate in vitro native lung cell–ECM crosstalk. This is a valuable system that can be easily applied to other organs for ECM-related drug screening, disease modeling, and basic mechanistic studies.
ER  -

GARLÍKOVÁ, Zuzana, Ana Catarina SILVA, Anas RABATA, David POTĚŠIL, Ivana IHNATOVÁ, Jana DUMKOVÁ, Zuzana KOLEDOVÁ, Zbyněk ZDRÁHAL, Vladimír VINARSKÝ, Aleš HAMPL, Perpétua PINTO-DO-Ó and Diana Santos NASCIMENTO. Generation of a Close-to-Native In Vitro System to Study Lung Cells-Extracellular Matrix Crosstalk. \textit{TISSUE ENGINEERING PART C-METHODS}. New Rochelle: Mary Ann Liebert, 2018, vol.~24, No~1, p.~1-13. ISSN~1937-3384. Available from: https://dx.doi.org/10.1089/ten.tec.2017.0283.

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