A new agarose-based microsystem to investigate cell response to prolonged confinement
| Autor: | Catherine Barentin, Sylvain Lefort, Charlotte Rivière, Véronique Maguer-Satta, Gilles Simon, Audrey Prunet, Jean-Paul Rieu, Hélène Delanoë-Ayari, Boris Guyot, S. Saci, B. Laperrousaz, Françoise Argoul, Stéphanie Gobert |
|---|---|
| Přispěvatelé: | Biophysique (BIOPHYSIQUE), Institut Lumière Matière [Villeurbanne] (ILM), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Liquides et interfaces (L&I), Laboratoire Ondes et Matière d'Aquitaine (LOMA), Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie Tissulaire et d'ingénierie Thérapeutique UMR 5305 (LBTI), Institut Universitaire de France (IUF), Ministère de l'Education nationale, de l’Enseignement supérieur et de la Recherche (M.E.N.E.S.R.), Laboratoire de Physique de l'ENS Lyon (Phys-ENS), École normale supérieure - Lyon (ENS Lyon)-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), This work was supported by the Institut Convergence PLASCAN, the Ligue Contre le Cancer foundation, the 'Institut Universitaire de France' (IUF), the 'Fondation pour la Recherche Médicale' (FRM) for part of A. Prunet's salary and the 'Fondation de France' and Alte SMP for part of S. Lefort's salary., VIDAL, Armelle |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Stromal cell [SDV.BIO]Life Sciences [q-bio]/Biotechnology [SPI] Engineering Sciences [physics] [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph] [SDV]Life Sciences [q-bio] Cell Biomedical Engineering Bioengineering [SDV.CAN]Life Sciences [q-bio]/Cancer 02 engineering and technology Matrix (biology) Biochemistry [PHYS] Physics [physics] 03 medical and health sciences [SPI]Engineering Sciences [physics] [CHIM] Chemical Sciences medicine [CHIM]Chemical Sciences Viability assay Cytoskeleton Cell Proliferation Cell Nucleus [PHYS]Physics [physics] [PHYS.PHYS.PHYS-OPTICS]Physics [physics]/Physics [physics]/Optics [physics.optics] Cell growth Sepharose [SPI.MECA.BIOM]Engineering Sciences [physics]/Mechanics [physics.med-ph]/Biomechanics [physics.med-ph] General Chemistry 021001 nanoscience & nanotechnology Cell biology Extracellular Matrix 030104 developmental biology medicine.anatomical_structure Cell culture 0210 nano-technology [PHYS.COND.CM-SCM]Physics [physics]/Condensed Matter [cond-mat]/Soft Condensed Matter [cond-mat.soft] Immunostaining |
| Zdroj: | Lab on a Chip Lab on a Chip, Royal Society of Chemistry, 2020, 20 (21), pp.4016-4030. ⟨10.1039/d0lc00732c⟩ Lab on a Chip, Royal Society of Chemistry, 2020, 20, pp.4016-4030. ⟨10.1039/D0LC00732C⟩ Lab on a Chip, 2020, 20, pp.4016-4030. ⟨10.1039/d1lc00192b⟩ Lab on a Chip, 2020, 20, pp.4016-4030. ⟨10.1039/D0LC00732C⟩ |
| ISSN: | 1473-0197 1473-0189 |
| Popis: | International audience; Emerging evidence suggests the importance of mechanical stimuli in normal and pathological situations for the control of many critical cellular functions. While the effect of matrix stiffness has been and is still extensively studied, few studies have focused on the role of mechanical stresses. The main limitation of such analyses is the lack of standard in vitro assays enabling extended mechanical stimulation compatible with dynamic biological and biophysical cell characterization. We have developed an agarose-based microsystem, the soft cell confiner, which enables the precise control of confinement for single or mixed cell populations. The rigidity of the confiner matches physiological conditions and its porosity enables passive medium renewal. It is compatible with time-lapse microscopy, in situ immunostaining, and standard molecular analyses, and can be used with both adherent and non-adherent cell lines. Cell proliferation of various cell lines (hematopoietic cells, MCF10A epithelial breast cells and HS27A stromal cells) was followed for several days up to confluence using video-microscopy and further documented by Western blot and immunostaining. Interestingly, even though the nuclear projected area was much larger upon confinement, with many highly deformed nuclei (non-circular shape), cell viability, assessed by live and dead cell staining, was unaffected for up to 8 days in the confiner. However, there was a decrease in cell proliferation upon confinement for all cell lines tested. The soft cell confiner is thus a valuable tool to decipher the effects of long-term confinement and deformation on the biology of cell populations. This tool will be instrumental in deciphering the impact of nuclear and cytoskeletal mechanosensitivity in normal and pathological conditions involving highly confined situations, such as those reported upon aging with fibrosis or during cancer. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|