Cargando…

A novel spheroid-based co-culture model mimics loss of keratinocyte differentiation, melanoma cell invasion, and drug-induced selection of ABCB5-expressing cells

BACKGROUND: Different 3D-cell culture approaches with varying degrees of complexity have been developed to serve as melanoma models for drug testing or mechanistic studies. While these 3D-culture initiatives are already often superior to classical 2D approaches, they are either composed of only mela...

Descripción completa

Detalles Bibliográficos
Autores principales: Klicks, Julia, Maßlo, Christoph, Kluth, Andreas, Rudolf, Rüdiger, Hafner, Mathias
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6489189/
https://www.ncbi.nlm.nih.gov/pubmed/31035967
http://dx.doi.org/10.1186/s12885-019-5606-4
_version_ 1783414769766629376
author Klicks, Julia
Maßlo, Christoph
Kluth, Andreas
Rudolf, Rüdiger
Hafner, Mathias
author_facet Klicks, Julia
Maßlo, Christoph
Kluth, Andreas
Rudolf, Rüdiger
Hafner, Mathias
author_sort Klicks, Julia
collection PubMed
description BACKGROUND: Different 3D-cell culture approaches with varying degrees of complexity have been developed to serve as melanoma models for drug testing or mechanistic studies. While these 3D-culture initiatives are already often superior to classical 2D approaches, they are either composed of only melanoma cells or they are so complex that the behavior of individual cell types is hard to understand, and often they are difficult to establish and expensive. METHODS: This study used low-attachment based generation of spheroids composed of up to three cell types. Characterization of cells and spheroids involved cryosectioning, immunofluorescence, FACS, and quantitative analyses. Statistical evaluation used one-way ANOVA with post-hoc Tukey test or Student’s t-test. RESULTS: The tri-culture model allowed to track cellular behavior in a cell-type specific manner and recapitulated different characteristics of early melanoma stages. Cells arranged into a collagen-IV rich fibroblast core, a ring of keratinocytes, and groups of highly proliferating melanoma cells on the outside. Regularly, some melanoma cells were also found to invade the fibroblast core. In the absence of melanoma cells, the keratinocyte ring stratified into central basal-like and peripheral, more differentiated cells. Conversely, keratinocyte differentiation was clearly reduced upon addition of melanoma cells. Treatment with the cytostatic drug, docetaxel, restored keratinocyte differentiation and induced apoptosis of external melanoma cells. Remaining intact external melanoma cells showed a significantly increased amount of ABCB5-immunoreactivity. CONCLUSIONS: In the present work, a novel, simple spheroid-based melanoma tri-culture model composed of fibroblasts, keratinocytes, and melanoma cells was described. This model mimicked features observed in early melanoma stages, including loss of keratinocyte differentiation, melanoma cell invasion, and drug-induced increase of ABCB5 expression in external melanoma cells. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12885-019-5606-4) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-6489189
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-64891892019-06-05 A novel spheroid-based co-culture model mimics loss of keratinocyte differentiation, melanoma cell invasion, and drug-induced selection of ABCB5-expressing cells Klicks, Julia Maßlo, Christoph Kluth, Andreas Rudolf, Rüdiger Hafner, Mathias BMC Cancer Research Article BACKGROUND: Different 3D-cell culture approaches with varying degrees of complexity have been developed to serve as melanoma models for drug testing or mechanistic studies. While these 3D-culture initiatives are already often superior to classical 2D approaches, they are either composed of only melanoma cells or they are so complex that the behavior of individual cell types is hard to understand, and often they are difficult to establish and expensive. METHODS: This study used low-attachment based generation of spheroids composed of up to three cell types. Characterization of cells and spheroids involved cryosectioning, immunofluorescence, FACS, and quantitative analyses. Statistical evaluation used one-way ANOVA with post-hoc Tukey test or Student’s t-test. RESULTS: The tri-culture model allowed to track cellular behavior in a cell-type specific manner and recapitulated different characteristics of early melanoma stages. Cells arranged into a collagen-IV rich fibroblast core, a ring of keratinocytes, and groups of highly proliferating melanoma cells on the outside. Regularly, some melanoma cells were also found to invade the fibroblast core. In the absence of melanoma cells, the keratinocyte ring stratified into central basal-like and peripheral, more differentiated cells. Conversely, keratinocyte differentiation was clearly reduced upon addition of melanoma cells. Treatment with the cytostatic drug, docetaxel, restored keratinocyte differentiation and induced apoptosis of external melanoma cells. Remaining intact external melanoma cells showed a significantly increased amount of ABCB5-immunoreactivity. CONCLUSIONS: In the present work, a novel, simple spheroid-based melanoma tri-culture model composed of fibroblasts, keratinocytes, and melanoma cells was described. This model mimicked features observed in early melanoma stages, including loss of keratinocyte differentiation, melanoma cell invasion, and drug-induced increase of ABCB5 expression in external melanoma cells. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12885-019-5606-4) contains supplementary material, which is available to authorized users. BioMed Central 2019-04-29 /pmc/articles/PMC6489189/ /pubmed/31035967 http://dx.doi.org/10.1186/s12885-019-5606-4 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Klicks, Julia
Maßlo, Christoph
Kluth, Andreas
Rudolf, Rüdiger
Hafner, Mathias
A novel spheroid-based co-culture model mimics loss of keratinocyte differentiation, melanoma cell invasion, and drug-induced selection of ABCB5-expressing cells
title A novel spheroid-based co-culture model mimics loss of keratinocyte differentiation, melanoma cell invasion, and drug-induced selection of ABCB5-expressing cells
title_full A novel spheroid-based co-culture model mimics loss of keratinocyte differentiation, melanoma cell invasion, and drug-induced selection of ABCB5-expressing cells
title_fullStr A novel spheroid-based co-culture model mimics loss of keratinocyte differentiation, melanoma cell invasion, and drug-induced selection of ABCB5-expressing cells
title_full_unstemmed A novel spheroid-based co-culture model mimics loss of keratinocyte differentiation, melanoma cell invasion, and drug-induced selection of ABCB5-expressing cells
title_short A novel spheroid-based co-culture model mimics loss of keratinocyte differentiation, melanoma cell invasion, and drug-induced selection of ABCB5-expressing cells
title_sort novel spheroid-based co-culture model mimics loss of keratinocyte differentiation, melanoma cell invasion, and drug-induced selection of abcb5-expressing cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6489189/
https://www.ncbi.nlm.nih.gov/pubmed/31035967
http://dx.doi.org/10.1186/s12885-019-5606-4
work_keys_str_mv AT klicksjulia anovelspheroidbasedcoculturemodelmimicslossofkeratinocytedifferentiationmelanomacellinvasionanddruginducedselectionofabcb5expressingcells
AT maßlochristoph anovelspheroidbasedcoculturemodelmimicslossofkeratinocytedifferentiationmelanomacellinvasionanddruginducedselectionofabcb5expressingcells
AT kluthandreas anovelspheroidbasedcoculturemodelmimicslossofkeratinocytedifferentiationmelanomacellinvasionanddruginducedselectionofabcb5expressingcells
AT rudolfrudiger anovelspheroidbasedcoculturemodelmimicslossofkeratinocytedifferentiationmelanomacellinvasionanddruginducedselectionofabcb5expressingcells
AT hafnermathias anovelspheroidbasedcoculturemodelmimicslossofkeratinocytedifferentiationmelanomacellinvasionanddruginducedselectionofabcb5expressingcells
AT klicksjulia novelspheroidbasedcoculturemodelmimicslossofkeratinocytedifferentiationmelanomacellinvasionanddruginducedselectionofabcb5expressingcells
AT maßlochristoph novelspheroidbasedcoculturemodelmimicslossofkeratinocytedifferentiationmelanomacellinvasionanddruginducedselectionofabcb5expressingcells
AT kluthandreas novelspheroidbasedcoculturemodelmimicslossofkeratinocytedifferentiationmelanomacellinvasionanddruginducedselectionofabcb5expressingcells
AT rudolfrudiger novelspheroidbasedcoculturemodelmimicslossofkeratinocytedifferentiationmelanomacellinvasionanddruginducedselectionofabcb5expressingcells
AT hafnermathias novelspheroidbasedcoculturemodelmimicslossofkeratinocytedifferentiationmelanomacellinvasionanddruginducedselectionofabcb5expressingcells