Planar AFM macro-probes to study the biomechanical properties of large cells and 3D cell spheroids

Andolfi, Laura; Greco, Silvio Mario Luciano; Tierno, Domenico; Chignola, Roberto; Martinelli, Monica; Giolo, Elena; Luppi, Stefania; Delfino, Ines; Zanetti, Michele; Battistella, Alice; Baldini, Giovanna; Ricci, Giuseppe; Lazzarino, Marco

doi:10.1016/j.actbio.2019.05.072

Cited by 32 publications

(28 citation statements)

References 46 publications

(66 reference statements)

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“…For WJ-MSCs, in the early stage of the cell spheroid growth, the surface pressure was high, resulting in high cell cohesion and aggregation. During the middle period to the late period of spheroid growth, the pressure was reduced, enabling rapid cell growth and amplification [42][43][44]. According to a previous study, increased pressure on the cell culture environment may promote the ossification of the MSCs [45].…”

Section: Discussionmentioning

confidence: 99%

A Dynamic Hanging-Drop System for Mesenchymal Stem Cell Culture

Huang

Tzeng

Chen

et al. 2020

IJMS

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There have been many microfluid technologies combined with hanging-drop for cell culture gotten developed in the past decade. A common problem within these devices is that the cell suspension introduced at the central inlet could cause a number of cells in each microwell to not regularize. Also, the instability of droplets during the spheroid formation remains an unsolved ordeal. In this study, we designed a microfluidic-based hanging-drop culture system with the design of taper-tube that can increase the stability of droplets while enhancing the rate of liquid exchange. A ring is surrounding the taper-tube. The ring can hold the cells to enable us to seed an adequate amount of cells before perfusion. Moreover, during the period of cell culture, the mechanical force around the cell is relatively low to prevent stem cells from differentiate and maintain the phenotype. As a result of our hanging system design, cells are designed to accumulate at the bottom of the droplet. This method enhances convenience for observation activities and analysis of experiments. Thus, this microfluid chip can be used as an in vitro platform representing in vivo physiological conditions, and can be useful in regenerative therapy.

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Section: Discussionmentioning

confidence: 99%

A Dynamic Hanging-Drop System for Mesenchymal Stem Cell Culture

Huang

Tzeng

Chen

et al. 2020

IJMS

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“…Stress-relaxation measurements were performed on human and mouse oocytes by lowering the z-piezo 20 µm beyond the contact point. In both cases, the analysis of the force-relaxation curves showed two distinct relaxation processes: a fast (1-2 s) and a slower relaxation (10-20 s) [69]. The presence of the two relaxation times suggests that the two main structural components of the oocyte, the ZP and the ooplasm, contribute differently to the mechanical response of the oocyte.…”

Section: The Mechanical Properties Of the Oocytementioning

confidence: 92%

Scanning Probe Microscopies: Imaging and Biomechanics in Reproductive Medicine Research

Andolfi

Battistella

Zanetti

et al. 2021

IJMS

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Basic and translational research in reproductive medicine can provide new insights with the application of scanning probe microscopies, such as atomic force microscopy (AFM) and scanning near-field optical microscopy (SNOM). These microscopies, which provide images with spatial resolution well beyond the optical resolution limit, enable users to achieve detailed descriptions of cell topography, inner cellular structure organization, and arrangements of single or cluster membrane proteins. A peculiar characteristic of AFM operating in force spectroscopy mode is its inherent ability to measure the interaction forces between single proteins or cells, and to quantify the mechanical properties (i.e., elasticity, viscoelasticity, and viscosity) of cells and tissues. The knowledge of the cell ultrastructure, the macromolecule organization, the protein dynamics, the investigation of biological interaction forces, and the quantification of biomechanical features can be essential clues for identifying the molecular mechanisms that govern responses in living cells. This review highlights the main findings achieved by the use of AFM and SNOM in assisted reproductive research, such as the description of gamete morphology; the quantification of mechanical properties of gametes; the role of forces in embryo development; the significance of investigating single-molecule interaction forces; the characterization of disorders of the reproductive system; and the visualization of molecular organization. New perspectives of analysis opened up by applying these techniques and the translational impacts on reproductive medicine are discussed.

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“…[ 148–150 ] Cantilever stiffness as well as the size and shape of the probe tip (affecting the contact area) can be adjusted to measure forces as low as several piconewtons and has been used to approximate the elastic properties of individual protein molecules and polymer chains. [ 151,152 ] Since indentation methods often allow only for the surface of a material to be indented, this approach is often not suitable for in situ measurements. However, for samples where the materials of interest lie near the surface, highly resolved maps of material stiffness can be generated by iteratively indenting the sample throughout an area of interest.…”

Section: Measuring Cell and Tissue Mechanical Properties For Mechanotmentioning

confidence: 99%

Biofabrication Strategies and Engineered In Vitro Systems for Vascular Mechanobiology

Pradhan

Banda

Farino

et al. 2020

Adv Healthcare Materials

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The vascular system is integral for maintaining organ‐specific functions and homeostasis. Dysregulation in vascular architecture and function can lead to various chronic or acute disorders. Investigation of the role of the vascular system in health and disease has been accelerated through the development of tissue‐engineered constructs and microphysiological on‐chip platforms. These in vitro systems permit studies of biochemical regulation of vascular networks and parenchymal tissue and provide mechanistic insights into the biophysical and hemodynamic forces acting in organ‐specific niches. Detailed understanding of these forces and the mechanotransductory pathways involved is necessary to develop preventative and therapeutic strategies targeting the vascular system. This review describes vascular structure and function, the role of hemodynamic forces in maintaining vascular homeostasis, and measurement approaches for cell and tissue level mechanical properties influencing vascular phenomena. State‐of‐the‐art techniques for fabricating in vitro microvascular systems, with varying degrees of biological and engineering complexity, are summarized. Finally, the role of vascular mechanobiology in organ‐specific niches and pathophysiological states, and efforts to recapitulate these events using in vitro microphysiological systems, are explored. It is hoped that this review will help readers appreciate the important, but understudied, role of vascular‐parenchymal mechanotransduction in health and disease toward developing mechanotherapeutics for treatment strategies.

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Planar AFM macro-probes to study the biomechanical properties of large cells and 3D cell spheroids

Cited by 32 publications

References 46 publications

A Dynamic Hanging-Drop System for Mesenchymal Stem Cell Culture

A Dynamic Hanging-Drop System for Mesenchymal Stem Cell Culture

Scanning Probe Microscopies: Imaging and Biomechanics in Reproductive Medicine Research

Biofabrication Strategies and Engineered In Vitro Systems for Vascular Mechanobiology

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