Quantifying cellular forces and biomechanical properties by correlative micropillar traction force and Brillouin microscopy

Coppola, Stefano; Schmidt, Thomas; Ruocco, G.; Antonacci, Giuseppe

doi:10.1364/boe.10.002202

Cited by 19 publications

(11 citation statements)

References 57 publications

(41 reference statements)

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“…As this imaging-based strategy considers the structural aspects of sarcomere maturation, contractility data were not assessed. In this regard, traction force microscopy or the use of fluorescent micropillars can help to correlate sarcomere quality and contraction force, complementing our microscopic approach [ 33 – 36 ]. In addition, the formation of a mature sarcomere network encompasses isoform switching of myofibril proteins, including titin and myosin heavy chain protein, which should also be considered to fully address sarcomere maturation [ 7 ].…”

Section: Discussionmentioning

confidence: 99%

Monitoring the maturation of the sarcomere network: a super-resolution microscopy-based approach

Skorska

Johann

Chabanovska

et al. 2022

Cell. Mol. Life Sci.

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The in vitro generation of human cardiomyocytes derived from induced pluripotent stem cells (iPSC) is of great importance for cardiac disease modeling, drug-testing applications and for regenerative medicine. Despite the development of various cultivation strategies, a sufficiently high degree of maturation is still a decisive limiting factor for the successful application of these cardiac cells. The maturation process includes, among others, the proper formation of sarcomere structures, mediating the contraction of cardiomyocytes. To precisely monitor the maturation of the contractile machinery, we have established an imaging-based strategy that allows quantitative evaluation of important parameters, defining the quality of the sarcomere network. iPSC-derived cardiomyocytes were subjected to different culture conditions to improve sarcomere formation, including prolonged cultivation time and micro patterned surfaces. Fluorescent images of α-actinin were acquired using super-resolution microscopy. Subsequently, we determined cell morphology, sarcomere density, filament alignment, z-Disc thickness and sarcomere length of iPSC-derived cardiomyocytes. Cells from adult and neonatal heart tissue served as control. Our image analysis revealed a profound effect on sarcomere content and filament orientation when iPSC-derived cardiomyocytes were cultured on structured, line-shaped surfaces. Similarly, prolonged cultivation time had a beneficial effect on the structural maturation, leading to a more adult-like phenotype. Automatic evaluation of the sarcomere filaments by machine learning validated our data. Moreover, we successfully transferred this approach to skeletal muscle cells, showing an improved sarcomere formation cells over different differentiation periods. Overall, our image-based workflow can be used as a straight-forward tool to quantitatively estimate the structural maturation of contractile cells. As such, it can support the establishment of novel differentiation protocols to enhance sarcomere formation and maturity.

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Section: Discussionmentioning

confidence: 99%

Monitoring the maturation of the sarcomere network: a super-resolution microscopy-based approach

Skorska

Johann

Chabanovska

et al. 2022

Cell. Mol. Life Sci.

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“…Brillouin microscopy is a method used to measure stiffness and viscoelastic properties through the material’s p-wave modulus in the GHz frequency range [ 105 , 106 ]. This non-contact 3D method of measurement works on the premise of photons (e.g., from a laser source) exciting microscopic acoustic waves in the material of interest ( Figure 2 ).…”

Section: Ex Vivo Assessment Of Pdac Biomechanicsmentioning

confidence: 99%

Pancreatic Ductal Adenocarcinoma: Relating Biomechanics and Prognosis

MacCurtain

Quirke

Thorpe

et al. 2021

JCM

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Pancreatic ductal adenocarcinoma (PDAC) is the most common form of pancreatic cancer and carries a dismal prognosis. Resectable patients are treated predominantly with surgery while borderline resectable patients may receive neoadjuvant treatment (NAT) to downstage their disease prior to possible resection. PDAC tissue is stiffer than healthy pancreas, and tissue stiffness is associated with cancer progression. Another feature of PDAC is increased tissue heterogeneity. We postulate that tumour stiffness and heterogeneity may be used alongside currently employed diagnostics to better predict prognosis and response to treatment. In this review we summarise the biomechanical changes observed in PDAC, explore the factors behind these changes and describe the clinical consequences. We identify methods available for assessing PDAC biomechanics ex vivo and in vivo, outlining the relative merits of each. Finally, we discuss the potential use of radiological imaging for prognostic use.

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“…The first category of techniques focuses on measuring the forces a cell can exert on its environment ( Figure 2 A): This is the case for traction force microscopy (TFM), where forces are inferred from the motion of probes embedded in the soft 2D substrate or 3D surrounding matrix [ 142 ], and of micropillar arrays, where the deflection of elastic pillars on the substrate that the cell adheres to indicates the amplitude of the cell traction force [ 143 , 144 ]. Such experiments, commonly used to study cell adhesion and migration, allow the establishment of the local force field and deciphering of the spatiotemporal coordination of the cell.…”

Section: Microfluidic Tools To Investigate Alterations Of the Cellmentioning

confidence: 99%

Molecular and Mechanobiological Pathways Related to the Physiopathology of FPLD2

Varlet

Helfer

Badens

2020

Cells

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Laminopathies are rare and heterogeneous diseases affecting one to almost all tissues, as in Progeria, and sharing certain features such as metabolic disorders and a predisposition to atherosclerotic cardiovascular diseases. These two features are the main characteristics of the adipose tissue-specific laminopathy called familial partial lipodystrophy type 2 (FPLD2). The only gene that is involved in FPLD2 physiopathology is the LMNA gene, with at least 20 mutations that are considered pathogenic. LMNA encodes the type V intermediate filament lamin A/C, which is incorporated into the lamina meshwork lining the inner membrane of the nuclear envelope. Lamin A/C is involved in the regulation of cellular mechanical properties through the control of nuclear rigidity and deformability, gene modulation and chromatin organization. While recent studies have described new potential signaling pathways dependent on lamin A/C and associated with FPLD2 physiopathology, the whole picture of how the syndrome develops remains unknown. In this review, we summarize the signaling pathways involving lamin A/C that are associated with the progression of FPLD2. We also explore the links between alterations of the cellular mechanical properties and FPLD2 physiopathology. Finally, we introduce potential tools based on the exploration of cellular mechanical properties that could be redirected for FPLD2 diagnosis.

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Quantifying cellular forces and biomechanical properties by correlative micropillar traction force and Brillouin microscopy

Cited by 19 publications

References 57 publications

Monitoring the maturation of the sarcomere network: a super-resolution microscopy-based approach

Monitoring the maturation of the sarcomere network: a super-resolution microscopy-based approach

Pancreatic Ductal Adenocarcinoma: Relating Biomechanics and Prognosis

Molecular and Mechanobiological Pathways Related to the Physiopathology of FPLD2

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