Fibroblasts maintained in 3 dimensions show a better differentiation state and higher sensitivity to estrogens

Claudia Montani, Nathalie Steimberg, Jennifer Boniotti, Giorgio Biasiotto, Isabella Zanella, Giuseppe Diafera, Ida Biunno, Luigi Caimi, Giovanna Mazzoleni, Diego Di Lorenzo

Research output: Contribution to journalArticle

Abstract

Cell differentiation and response to hormonal signals were studied in a 3D environment on an in-house generated mouse fibroblast cell line expressing a reporter gene under the control of estrogen responsive sequences (EREs). 3D cell culture conditions were obtained in a Rotary Cell Culture System; (RCCS™), a microgravity based bioreactor that promotes the aggregation of cells into multicellular spheroids (MCS). In this bioreactor the cells maintained a better differentiated phenotype and more closely resembled in vivo tissue. The RCCS™ cultured fibroblasts showed higher expression of genes regulating cell assembly, differentiation and hormonal functions. Microarray analysis showed that genes related to cell cycle, proliferation, cytoskeleton, migration, adhesion and motility were all down-regulated in 3D as compared to 2D conditions, as well as oncogene expression and inflammatory cytokines. Controlled remodeling of ECM, which is an essential aspect of cell organization, homeostasis and tissue was affected by the culture method as assessed by immunolocalization of β-tubulin. Markers of cell organization, homeostasis and tissue repair, metalloproteinase 2 (MMP2) and its physiological inhibitor (TIMP4) changed expression in association with the relative formation of cell aggregates. The fibroblasts cultured in the RCCS™ maintain a better responsiveness to estrogens, measured as expression of ERα and regulation of an ERE-dependent reporter and of the endogenous target genes CBP, Rarb, MMP1 and Dbp.Our data highlight the interest of this 3D culture model for its potential application in the field of cell response to hormonal signals and the pharmaco-toxicological analyses of chemicals and natural molecules endowed of estrogenic potential.

Original languageEnglish
Pages (from-to)421-433
Number of pages13
JournalToxicology and Applied Pharmacology
Volume280
Issue number3
DOIs
Publication statusPublished - Nov 1 2014

Fingerprint

Fibroblasts
Estrogens
Genes
Cells
Tissue
Bioreactors
Cell culture
Cell Differentiation
Military electronic countermeasures
Homeostasis
Cell Culture Techniques
Cellular Spheroids
Microgravity
Metalloproteases
Tubulin
Microarrays
Weightlessness
Cell Aggregation
Matrix Metalloproteinase 2
Microarray Analysis

Keywords

  • 3 dimension
  • Bioreactor
  • ECM
  • Estrogen receptor
  • Fibroblasts
  • Microgravity

ASJC Scopus subject areas

  • Pharmacology
  • Toxicology
  • Medicine(all)

Cite this

Montani, C., Steimberg, N., Boniotti, J., Biasiotto, G., Zanella, I., Diafera, G., ... Di Lorenzo, D. (2014). Fibroblasts maintained in 3 dimensions show a better differentiation state and higher sensitivity to estrogens. Toxicology and Applied Pharmacology, 280(3), 421-433. https://doi.org/10.1016/j.taap.2014.08.021

Fibroblasts maintained in 3 dimensions show a better differentiation state and higher sensitivity to estrogens. / Montani, Claudia; Steimberg, Nathalie; Boniotti, Jennifer; Biasiotto, Giorgio; Zanella, Isabella; Diafera, Giuseppe; Biunno, Ida; Caimi, Luigi; Mazzoleni, Giovanna; Di Lorenzo, Diego.

In: Toxicology and Applied Pharmacology, Vol. 280, No. 3, 01.11.2014, p. 421-433.

Research output: Contribution to journalArticle

Montani, C, Steimberg, N, Boniotti, J, Biasiotto, G, Zanella, I, Diafera, G, Biunno, I, Caimi, L, Mazzoleni, G & Di Lorenzo, D 2014, 'Fibroblasts maintained in 3 dimensions show a better differentiation state and higher sensitivity to estrogens', Toxicology and Applied Pharmacology, vol. 280, no. 3, pp. 421-433. https://doi.org/10.1016/j.taap.2014.08.021
Montani, Claudia ; Steimberg, Nathalie ; Boniotti, Jennifer ; Biasiotto, Giorgio ; Zanella, Isabella ; Diafera, Giuseppe ; Biunno, Ida ; Caimi, Luigi ; Mazzoleni, Giovanna ; Di Lorenzo, Diego. / Fibroblasts maintained in 3 dimensions show a better differentiation state and higher sensitivity to estrogens. In: Toxicology and Applied Pharmacology. 2014 ; Vol. 280, No. 3. pp. 421-433.
@article{3ad746c757324b2b8a4ae5e04ac05d64,
title = "Fibroblasts maintained in 3 dimensions show a better differentiation state and higher sensitivity to estrogens",
abstract = "Cell differentiation and response to hormonal signals were studied in a 3D environment on an in-house generated mouse fibroblast cell line expressing a reporter gene under the control of estrogen responsive sequences (EREs). 3D cell culture conditions were obtained in a Rotary Cell Culture System; (RCCS™), a microgravity based bioreactor that promotes the aggregation of cells into multicellular spheroids (MCS). In this bioreactor the cells maintained a better differentiated phenotype and more closely resembled in vivo tissue. The RCCS™ cultured fibroblasts showed higher expression of genes regulating cell assembly, differentiation and hormonal functions. Microarray analysis showed that genes related to cell cycle, proliferation, cytoskeleton, migration, adhesion and motility were all down-regulated in 3D as compared to 2D conditions, as well as oncogene expression and inflammatory cytokines. Controlled remodeling of ECM, which is an essential aspect of cell organization, homeostasis and tissue was affected by the culture method as assessed by immunolocalization of β-tubulin. Markers of cell organization, homeostasis and tissue repair, metalloproteinase 2 (MMP2) and its physiological inhibitor (TIMP4) changed expression in association with the relative formation of cell aggregates. The fibroblasts cultured in the RCCS™ maintain a better responsiveness to estrogens, measured as expression of ERα and regulation of an ERE-dependent reporter and of the endogenous target genes CBP, Rarb, MMP1 and Dbp.Our data highlight the interest of this 3D culture model for its potential application in the field of cell response to hormonal signals and the pharmaco-toxicological analyses of chemicals and natural molecules endowed of estrogenic potential.",
keywords = "3 dimension, Bioreactor, ECM, Estrogen receptor, Fibroblasts, Microgravity",
author = "Claudia Montani and Nathalie Steimberg and Jennifer Boniotti and Giorgio Biasiotto and Isabella Zanella and Giuseppe Diafera and Ida Biunno and Luigi Caimi and Giovanna Mazzoleni and {Di Lorenzo}, Diego",
year = "2014",
month = "11",
day = "1",
doi = "10.1016/j.taap.2014.08.021",
language = "English",
volume = "280",
pages = "421--433",
journal = "Toxicology and Applied Pharmacology",
issn = "0041-008X",
publisher = "Academic Press Inc.",
number = "3",

}

TY - JOUR

T1 - Fibroblasts maintained in 3 dimensions show a better differentiation state and higher sensitivity to estrogens

AU - Montani, Claudia

AU - Steimberg, Nathalie

AU - Boniotti, Jennifer

AU - Biasiotto, Giorgio

AU - Zanella, Isabella

AU - Diafera, Giuseppe

AU - Biunno, Ida

AU - Caimi, Luigi

AU - Mazzoleni, Giovanna

AU - Di Lorenzo, Diego

PY - 2014/11/1

Y1 - 2014/11/1

N2 - Cell differentiation and response to hormonal signals were studied in a 3D environment on an in-house generated mouse fibroblast cell line expressing a reporter gene under the control of estrogen responsive sequences (EREs). 3D cell culture conditions were obtained in a Rotary Cell Culture System; (RCCS™), a microgravity based bioreactor that promotes the aggregation of cells into multicellular spheroids (MCS). In this bioreactor the cells maintained a better differentiated phenotype and more closely resembled in vivo tissue. The RCCS™ cultured fibroblasts showed higher expression of genes regulating cell assembly, differentiation and hormonal functions. Microarray analysis showed that genes related to cell cycle, proliferation, cytoskeleton, migration, adhesion and motility were all down-regulated in 3D as compared to 2D conditions, as well as oncogene expression and inflammatory cytokines. Controlled remodeling of ECM, which is an essential aspect of cell organization, homeostasis and tissue was affected by the culture method as assessed by immunolocalization of β-tubulin. Markers of cell organization, homeostasis and tissue repair, metalloproteinase 2 (MMP2) and its physiological inhibitor (TIMP4) changed expression in association with the relative formation of cell aggregates. The fibroblasts cultured in the RCCS™ maintain a better responsiveness to estrogens, measured as expression of ERα and regulation of an ERE-dependent reporter and of the endogenous target genes CBP, Rarb, MMP1 and Dbp.Our data highlight the interest of this 3D culture model for its potential application in the field of cell response to hormonal signals and the pharmaco-toxicological analyses of chemicals and natural molecules endowed of estrogenic potential.

AB - Cell differentiation and response to hormonal signals were studied in a 3D environment on an in-house generated mouse fibroblast cell line expressing a reporter gene under the control of estrogen responsive sequences (EREs). 3D cell culture conditions were obtained in a Rotary Cell Culture System; (RCCS™), a microgravity based bioreactor that promotes the aggregation of cells into multicellular spheroids (MCS). In this bioreactor the cells maintained a better differentiated phenotype and more closely resembled in vivo tissue. The RCCS™ cultured fibroblasts showed higher expression of genes regulating cell assembly, differentiation and hormonal functions. Microarray analysis showed that genes related to cell cycle, proliferation, cytoskeleton, migration, adhesion and motility were all down-regulated in 3D as compared to 2D conditions, as well as oncogene expression and inflammatory cytokines. Controlled remodeling of ECM, which is an essential aspect of cell organization, homeostasis and tissue was affected by the culture method as assessed by immunolocalization of β-tubulin. Markers of cell organization, homeostasis and tissue repair, metalloproteinase 2 (MMP2) and its physiological inhibitor (TIMP4) changed expression in association with the relative formation of cell aggregates. The fibroblasts cultured in the RCCS™ maintain a better responsiveness to estrogens, measured as expression of ERα and regulation of an ERE-dependent reporter and of the endogenous target genes CBP, Rarb, MMP1 and Dbp.Our data highlight the interest of this 3D culture model for its potential application in the field of cell response to hormonal signals and the pharmaco-toxicological analyses of chemicals and natural molecules endowed of estrogenic potential.

KW - 3 dimension

KW - Bioreactor

KW - ECM

KW - Estrogen receptor

KW - Fibroblasts

KW - Microgravity

UR - http://www.scopus.com/inward/record.url?scp=84910070339&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84910070339&partnerID=8YFLogxK

U2 - 10.1016/j.taap.2014.08.021

DO - 10.1016/j.taap.2014.08.021

M3 - Article

C2 - 25173957

AN - SCOPUS:84910070339

VL - 280

SP - 421

EP - 433

JO - Toxicology and Applied Pharmacology

JF - Toxicology and Applied Pharmacology

SN - 0041-008X

IS - 3

ER -