<i>Ex vivo</i>live cell tracking in kidney organoids using light sheet fluorescence microscopy

Held, Marie; Santeramo, Ilaria; Wilm, Bettina; Murray, Patricia; Lévy, Raphaël

doi:10.1101/233114

Cited by 2 publications

(1 citation statement)

References 53 publications

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“…Additionally, this printing method can be combined with other organoid fabrication methods [32] and could provide a way to mass-produce organoids highly reproducible in size and shape. Combined with organoid microinjection [45] and advanced imaging methods [46], organoids could potentially replace animal models as a screening tool for drug testing, thereby reducing the research costs for new drugs.…”

Section: Discussionmentioning

confidence: 99%

Microfluidic-assisted bioprinting of tissues and organoids at high cell concentrations

et al. 2021

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Despite its simplicity, which makes it the most commonly used bioprinting method today, extrusion-based bioprinting suffers from its inability to reproduce the complex tissue architecture found in organs. Generally, this printing method allows for the dispensing of solutions of a predefined cell concentration through a rudimentary needle. Moreover, to avoid cell lysis in the dispensing needle, which is detrimental to the viability of the printed tissue, as well as cell loss in dead volumes of tubing, thereby increasing the cost of printing tissue, a common strategy has been to print with cell concentrations much lower in comparison to the concentrations found in living tissues. As a result, cell-to-cell distance is increased in the dispensed samples impairing communication through cytokines. Here, we present a microfluidic-based print head capable of modulating the printed cell concentration in real-time. This device allows bioprinting at high cell concentrations by concentrating and dispensing fibroblasts at concentrations up to 10 million cells∙mL−1. We also demonstrate that this device can be used to print bladder organoids. As the cell seeding concentration is of major importance for organogenesis in 3D culture, organoid printing allows the user to standardize the process of organoid formation and achieve more reliable and reproducible results.

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Section: Discussionmentioning

confidence: 99%

Microfluidic-assisted bioprinting of tissues and organoids at high cell concentrations

et al. 2021

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Novel Microscopic Techniques for Podocyte Research

Siegerist

Endlich

2018

Front. Endocrinol.

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Together with endothelial cells and the glomerular basement membrane, podocytes form the size-specific filtration barrier of the glomerulus with their interdigitating foot processes. Since glomerulopathies are associated with so-called foot process effacement—a severe change of well-formed foot processes into flat and broadened processes—visualization of the three-dimensional podocyte morphology is a crucial part for diagnosis of nephrotic diseases. However, interdigitating podocyte foot processes are too narrow to be resolved by classic light microscopy due to Ernst Abbe's law making electron microscopy necessary. Although three dimensional electron microscopy approaches like serial block face and focused ion beam scanning electron microscopy and electron tomography allow volumetric reconstruction of podocytes, these techniques are very time-consuming and too specialized for routine use or screening purposes. During the last few years, different super-resolution microscopic techniques were developed to overcome the optical resolution limit enabling new insights into podocyte morphology. Super-resolution microscopy approaches like three dimensional structured illumination microscopy (3D-SIM), stimulated emission depletion microscopy (STED) and localization microscopy [stochastic optical reconstruction microscopy (STORM), photoactivated localization microscopy (PALM)] reach resolutions down to 80–20 nm and can be used to image and further quantify podocyte foot process morphology. Furthermore, in vivo imaging of podocytes is essential to study the behavior of these cells in situ. Therefore, multiphoton laser microscopy was a breakthrough for in vivo studies of podocytes in transgenic animal models like rodents and zebrafish larvae because it allows imaging structures up to several hundred micrometer in depth within the tissue. Additionally, along with multiphoton microscopy, lightsheet microscopy is currently used to visualize larger tissue volumes and therefore image complete glomeruli in their native tissue context. Alongside plain visualization of cellular structures, atomic force microscopy has been used to study the change of mechanical properties of podocytes in diseased states which has been shown to be a culprit in podocyte maintenance. This review discusses recent advances in the field of microscopic imaging and demonstrates their currently used and other possible applications for podocyte research.

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Ex vivolive cell tracking in kidney organoids using light sheet fluorescence microscopy

Cited by 2 publications

References 53 publications

Microfluidic-assisted bioprinting of tissues and organoids at high cell concentrations

Microfluidic-assisted bioprinting of tissues and organoids at high cell concentrations

Novel Microscopic Techniques for Podocyte Research

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