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Impact of the composition and stiffness of a 3D hyaluronic acid-based
hydroscaffold on breast cancer cell growth and CD44 ...
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Impact of the composition and stiffness of a 3D hyaluronic acid-based hydroscaffold on breast cancer cell growth and CD44 expression

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The extracellular matrix (ECM) is present in all solid tissues and is a master regulator of cell behavior and phenotype. The ECM in each tissue is characterized by its biochemical and biophysical properties, which are modified in many different types of disease, especially cancer. Hyaluronic acid (HA), as a biopolymer, has emerged as a remarkable ECM component by its viscoelastic properties, its hygroscopic capacities and the diversity of cell processes it controls in health and disease.
BIOMIMESYS®, a patented HA-based hydroscaffold, is a highly reproducible and physiological matrix that is suitable for long-term 3D cell culture. Using BIOMIMESYS® Oncology - made of HA biofunctionalized with undenaturated collagen I - we investigated the influence of (i) matrix composition (with/without laminin) and (ii) stiffness on the growth and expression of CD44 (a receptor of HA) in spheroids made of breast cancer cell lines (MCF7, MDA-MB-231) and normal cells (MCF10A) along time. Analyses were carried out using high content analysis/screening (HCA/HCS, high throughput confocal microscopy).
As a whole, the results showed that the cell growth was lowered in the presence of laminin but promoted with increased stiffness. Except for MCF10A cells, the expression of CD44 had the tendency to increase over time, while the stiffness and the presence of laminin had no significant impact on its expression. Given that BIOMIMESYS® is a highly defined and easily tunable matrix, this physiological 3D cell culture system allows to study the impact of biochemical and biophysical properties of the matrix.
By better mimicking the cancer microenvironment - including the ECM - the new generation of 3D cell culture systems might help to discover new effective anti-cancer therapies. In this frame, our aim is to study the response of cancer cells in BIOMIMESYS® to a large set of anti-tumor molecules, using our HCS platform.
Keywords: 3D cell culture, hyaluronic acid, breast cancer, stiffness, high content screening

Publicado en: Salud y medicina
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Impact of the composition and stiffness of a 3D hyaluronic acid-based hydroscaffold on breast cancer cell growth and CD44 expression

  1. 1. Impact of the composition and stiffness of a 3D hyaluronic acid-based hydroscaffold on breast cancer cell growth and CD44 expression Abstract The extracellular matrix (ECM) is present in all solid tissues and is a master regulator of cell behavior and phenotype. The ECM in each tissue is characterized by its biochemical and biophysical properties, which are modified in many different types of disease, especially cancer. Hyaluronic acid (HA), as a biopolymer, has emerged as a remarkable ECM component by its viscoelastic properties, its hygroscopic capacities and the diversity of cell processes it controls in health and disease. BIOMIMESYS®, a patented HA-based hydroscaffold, is a highly reproducible and physiological matrix that is suitable for long-term 3D cell culture. Using BIOMIMESYS® Oncology - made of HA biofunctionalized with undenaturated collagen I - we investigated the influence of (i) matrix composition (with/without laminin) and (ii) stiffness on the growth and expression of CD44 (a receptor of HA) in spheroids made of breast cancer cell lines (MCF7, MDA-MB-231) and normal cells (MCF10A) along time. Analyses were carried out using high content analysis/screening (HCA/HCS, high throughput confocal microscopy). As a whole, the results showed that the cell growth was lowered in the presence of laminin but promoted with increased stiffness. Except for MCF10A cells, the expression of CD44 had the tendency to increase over time, while the stiffness and the presence of laminin had no significant impact on its expression. Given that BIOMIMESYS® is a highly defined and easily tunable matrix, this physiological 3D cell culture system allows to study the impact of biochemical and biophysical properties of the matrix. By better mimicking the cancer microenvironment - including the ECM - the new generation of 3D cell culture systems might help to discover new effective anti-cancer therapies. In this frame, our aim is to study the response of cancer cells in BIOMIMESYS® to a large set of anti-tumor molecules, using our HCS platform. Keywords: 3D cell culture, hyaluronic acid, breast cancer, stiffness, high content screening Methods Results Conclusions and perspectives Marie Lesaffre1, Noémie Carlier1, Nathalie Maubon1, Zied Souguir1, Elodie Vandenhaute1 1HCS Pharma, Biocentre Fleming, 250 rue Salvador Allende, 59120 Loos, FRANCE • Cell culture in hydroscaffold 96-well plate with the lyophilised hydroscaffolds Scaffold visualized by SEM • Cell lines used • Timeline MCF10A “normal-like" breast epithelial cell line MCF7 luminal A breast cancer cell line MDA-MB-231 triple negative breast cancer cell line • What is high content analysis/screening? Source: https://www.nottingham.ac.uk/life-sciences/facilities/slim/cell-signalling-imaging/high-content-screening/index.aspx • Effect of matrix stiffness on cancer cell growth • Effect of laminin on cell growth and CD44 expression These preliminary results show that BIOMIMESYS® is a HA-based relevant matrix to study the impact of the ECM on cancer cell growth and expression of proteins in long-lasting in vitro studies (one month, which would be difficult - if not impossible - in 2D cultures). Compatible with all classical downstream analysis methods, the format of BIOMIMESYS® (96-well, and now 384-well format) can also be beneficial in the frame of phenotypic screening in a search for new antitumor therapies. As such, our aim is to use BIOMIMESYS® as a tool for finding efficient molecules against each cancer type, by better mimicking the structure, the composition and the physico-chemical properties of the pathological ECM. Calcein green + Laminin MDA-MB-231 Day7 - Laminin (control) CD44 MDA-MB-231 Day14 The growth of spheroids seems to be lowered in the presence of laminin in BIOMIMESYS®.  The expression of CD44 varies over time, but the presence of laminin in BIOMIMESYS® seems not to impact its expression… What about CD44 variants? 0 5 0 0 0 0 1 0 0 0 0 0 1 5 0 0 0 0 2 0 0 0 0 0 2 5 0 0 0 0 M D A -M B -2 3 1 T im e (d a y s ) Spheroidarea(µm 2 ) B IO M IM E S Y S ® O n c o lo g y so ft B IO M IM E S Y S ® O n c o lo g y rig id 7 14 21 28 0 5 0 0 0 0 1 0 0 0 0 0 1 5 0 0 0 0 2 0 0 0 0 0 2 5 0 0 0 0 M D A -M B -2 3 1 T im e (d a y s ) Spheroidarea(µm 2 ) B IO M IM E S Y S ® O n c o lo g y so ft B IO M IM E S Y S ® O n c o lo g y rig id 7 14 21 28 0 5 0 0 0 0 1 0 0 0 0 0 1 5 0 0 0 0 2 0 0 0 0 0 2 5 0 0 0 0 M C F 1 0 A T im e (d a y s ) Spheroidarea(µm 2 ) B IO M IM E S Y S ® O n c o lo g y so ft B IO M IM E S Y S ® O n c o lo g y rig id 7 14 21 28 ~ 1 kPa ~ 10 kPa as measured by rheology  The matrix stiffness can be modulated in BIOMIMESYS®, which will allow us to study in more detail the impact of this parameter in ‘microenvironmentally-relevant’ conditions.

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