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Dermal fibroblasts and triple-negative mammary epithelial cancer cells differentially stiffen their local matrix

The bulk measurement of extracellular matrix (ECM) stiffness is commonly used in mechanobiology. However, past studies by our group show that peri-cellular stiffness is quite heterogeneous and divergent from the bulk. We use optical tweezers active microrheology (AMR) to quantify how two phenotypica...

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Autores principales: Jagiełło, Alicja, Lim, Micah, Botvinick, Elliot
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AIP Publishing LLC 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7719046/
https://www.ncbi.nlm.nih.gov/pubmed/33305163
http://dx.doi.org/10.1063/5.0021030
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author Jagiełło, Alicja
Lim, Micah
Botvinick, Elliot
author_facet Jagiełło, Alicja
Lim, Micah
Botvinick, Elliot
author_sort Jagiełło, Alicja
collection PubMed
description The bulk measurement of extracellular matrix (ECM) stiffness is commonly used in mechanobiology. However, past studies by our group show that peri-cellular stiffness is quite heterogeneous and divergent from the bulk. We use optical tweezers active microrheology (AMR) to quantify how two phenotypically distinct migratory cell lines establish dissimilar patterns of peri-cellular stiffness. Dermal fibroblasts (DFs) and triple-negative human breast cancer cells MDA-MB-231 (MDAs) were embedded within type 1 collagen (T1C) hydrogels polymerized at two concentrations: 1.0 mg/ml and 1.5 mg/ml. We found DFs increase the local stiffness of 1.0 mg/ml T1C hydrogels but, surprisingly, do not alter the stiffness of 1.5 mg/ml T1C hydrogels. In contrast, MDAs predominantly do not stiffen T1C hydrogels as compared to cell-free controls. The results suggest that MDAs adapt to the bulk ECM stiffness, while DFs regulate local stiffness to levels they intrinsically prefer. In other experiments, cells were treated with transforming growth factor-β1 (TGF-β1), glucose, or ROCK inhibitor Y27632, which have known effects on DFs and MDAs related to migration, proliferation, and contractility. The results show that TGF-β1 alters stiffness anisotropy, while glucose increases stiffness magnitude around DFs but not MDAs and Y27632 treatment inhibits cell-mediated stiffening. Both cell lines exhibit an elongated morphology and local stiffness anisotropy, where the stiffer axis depends on the cell line, T1C concentration, and treatment. In summary, our findings demonstrate that AMR reveals otherwise masked mechanical properties such as spatial gradients and anisotropy, which are known to affect cell behavior at the macro-scale. The same properties manifest with similar magnitude around single cells.
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spelling pubmed-77190462020-12-09 Dermal fibroblasts and triple-negative mammary epithelial cancer cells differentially stiffen their local matrix Jagiełło, Alicja Lim, Micah Botvinick, Elliot APL Bioeng Articles The bulk measurement of extracellular matrix (ECM) stiffness is commonly used in mechanobiology. However, past studies by our group show that peri-cellular stiffness is quite heterogeneous and divergent from the bulk. We use optical tweezers active microrheology (AMR) to quantify how two phenotypically distinct migratory cell lines establish dissimilar patterns of peri-cellular stiffness. Dermal fibroblasts (DFs) and triple-negative human breast cancer cells MDA-MB-231 (MDAs) were embedded within type 1 collagen (T1C) hydrogels polymerized at two concentrations: 1.0 mg/ml and 1.5 mg/ml. We found DFs increase the local stiffness of 1.0 mg/ml T1C hydrogels but, surprisingly, do not alter the stiffness of 1.5 mg/ml T1C hydrogels. In contrast, MDAs predominantly do not stiffen T1C hydrogels as compared to cell-free controls. The results suggest that MDAs adapt to the bulk ECM stiffness, while DFs regulate local stiffness to levels they intrinsically prefer. In other experiments, cells were treated with transforming growth factor-β1 (TGF-β1), glucose, or ROCK inhibitor Y27632, which have known effects on DFs and MDAs related to migration, proliferation, and contractility. The results show that TGF-β1 alters stiffness anisotropy, while glucose increases stiffness magnitude around DFs but not MDAs and Y27632 treatment inhibits cell-mediated stiffening. Both cell lines exhibit an elongated morphology and local stiffness anisotropy, where the stiffer axis depends on the cell line, T1C concentration, and treatment. In summary, our findings demonstrate that AMR reveals otherwise masked mechanical properties such as spatial gradients and anisotropy, which are known to affect cell behavior at the macro-scale. The same properties manifest with similar magnitude around single cells. AIP Publishing LLC 2020-12-04 /pmc/articles/PMC7719046/ /pubmed/33305163 http://dx.doi.org/10.1063/5.0021030 Text en © 2020 Author(s). 2473-2877/2020/4(4)/046105/12 All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Articles
Jagiełło, Alicja
Lim, Micah
Botvinick, Elliot
Dermal fibroblasts and triple-negative mammary epithelial cancer cells differentially stiffen their local matrix
title Dermal fibroblasts and triple-negative mammary epithelial cancer cells differentially stiffen their local matrix
title_full Dermal fibroblasts and triple-negative mammary epithelial cancer cells differentially stiffen their local matrix
title_fullStr Dermal fibroblasts and triple-negative mammary epithelial cancer cells differentially stiffen their local matrix
title_full_unstemmed Dermal fibroblasts and triple-negative mammary epithelial cancer cells differentially stiffen their local matrix
title_short Dermal fibroblasts and triple-negative mammary epithelial cancer cells differentially stiffen their local matrix
title_sort dermal fibroblasts and triple-negative mammary epithelial cancer cells differentially stiffen their local matrix
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7719046/
https://www.ncbi.nlm.nih.gov/pubmed/33305163
http://dx.doi.org/10.1063/5.0021030
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AT botvinickelliot dermalfibroblastsandtriplenegativemammaryepithelialcancercellsdifferentiallystiffentheirlocalmatrix