A Novel Microgravity Simulator Applicable for Three-Dimensional Cell Culturing

Wüest, Simon L.; Richard, Stéphane; Walther, Isabelle; Furrer, Reinhard; Anderegg, Roland; Sekler, Jörg; Egli, Marcel

doi:10.1007/s12217-014-9364-2

Cited by 30 publications

(40 citation statements)

References 29 publications

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“…For RPM modes the force calculation is more difficult, because there are two frames rotated and the speed and direction is changed during the mode of operation. As an approximation we used 2.41, a number derived from calculation of the farthest position of the sample during rotation (see Wüest et al 2014) and assuming both frames rotate constantly with a maximum speed of 10 rpm (~1.05 rad/s) and a maximum radius from the center of rotation of 0.0175 m (length of cuvette is 3.5 cm): α ¼ 2:41*ω 2 * r ¼ ∼0:05 g achieved over time due to random changes in speed and direction ð Þ…”

Section: Force Calculationsmentioning

confidence: 99%

Validation of Random Positioning Versus Clinorotation Using a Macrophage Model System

Brungs

Hauslage

Hemmersbach

2019

Microgravity Sci. Technol.

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Clinostats and Random Positioning Machines (RPMs) are valuable devices for microgravity simulations in order to study fundamental gravity-dependent mechanisms on ground and in preparation for space flights. Both devices have different modes of operation, which have to be carefully considered and comprehensively discussed with respect to their potential impact on the quality of (simulated) microgravity. Here, we used the well-studied oxidative burst reaction of the immune cell (macrophage) model system, in order to compare clinorotation with random positioning. The RPM was used in a clinorotation mode, rotating the sample with 60 rpm around the horizontal axis and in the "random speed mode" (2-10 rpm) with and without random direction, thus, two axes rotating either bi-or unidirectionally. The production of Reactive Oxygen Species (ROS) during oxidative burst of the macrophages was visualized by the luminescence luminol assay. During clinorotation the cells responded with a reduction of the ROS production which is fully in line with earlier studies (clinostat, parabolic flight). In contrast, the exposure to the two random positioning modes showed that the oxidative burst response during RPM-exposure differs significantly from that observed during clinorotation, i.e. a jittering, more variant form of ROS production. We can conclude from our work, that the RPM is not suitable in its real random mode (with and without random direction; 2-10 rpm) to simulate the conditions of microgravity for the chosen system. We recommend that investigators using microgravity simulators should carefully choose the device and mode of operation specifically for their cellular system of interest.

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Section: Force Calculationsmentioning

confidence: 99%

Validation of Random Positioning Versus Clinorotation Using a Macrophage Model System

Brungs

Hauslage

Hemmersbach

2019

Microgravity Sci. Technol.

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“…We have recently reported another approach to upgrading the RPM by installing a commercial CO 2 incubator onto the rotating frames. This RPM, called “random positioning incubator” (RPI) [ 9 , 10 ], has the advantage of being independent of large laboratory incubators ( Figure 1 ). Furthermore, the closed chamber of the incubator isolates the environment of the culture flasks and thus prevents exposure of biological samples to vapor and wear from the machinery, for example, [ 10 ].…”

Section: Rpm Development and Technologymentioning

confidence: 99%

“…This RPM, called “random positioning incubator” (RPI) [ 9 , 10 ], has the advantage of being independent of large laboratory incubators ( Figure 1 ). Furthermore, the closed chamber of the incubator isolates the environment of the culture flasks and thus prevents exposure of biological samples to vapor and wear from the machinery, for example, [ 10 ]. Besides differences in the design of the three RPM types (regular RPM, small desktop RPM, and RPI), there appear to be slightly different concepts of how to average the gravity vector.…”

Section: Rpm Development and Technologymentioning

confidence: 99%

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Simulated Microgravity: Critical Review on the Use of Random Positioning Machines for Mammalian Cell Culture

Wüest

Richard

Kopp

et al. 2015

BioMed Research International

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180

172

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Random Positioning Machines (RPMs) have been used since many years as a ground-based model to simulate microgravity. In this review we discuss several aspects of the RPM. Recent technological development has expanded the operative range of the RPM substantially. New possibilities of live cell imaging and partial gravity simulations, for example, are of particular interest. For obtaining valuable and reliable results from RPM experiments, the appropriate use of the RPM is of utmost importance. The simulation of microgravity requires that the RPM's rotation is faster than the biological process under study, but not so fast that undesired side effects appear. It remains a legitimate question, however, whether the RPM can accurately and reliably simulate microgravity conditions comparable to real microgravity in space. We attempt to answer this question by mathematically analyzing the forces working on the samples while they are mounted on the operating RPM and by comparing data obtained under real microgravity in space and simulated microgravity on the RPM. In conclusion and after taking the mentioned constraints into consideration, we are convinced that simulated microgravity experiments on the RPM are a valid alternative for conducting examinations on the influence of the force of gravity in a fast and straightforward approach.

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“…The RPM consists of two gimbal-mounted frames independently moved by motors. Samples are placed at the centre of the frames, where the generated motion pattern is designed to equally distribute the gravity vector spatially and temporally (the gravity mathematically averages zero) [33]. Because the constant reorientation of the gravity vector prevents biological systems to adjust to this force, their response is similar to the one achieved upon real microgravity exposure [16].…”

Section: Simulated Microgravitymentioning

confidence: 99%

TRPC6 in simulated microgravity of intervertebral disc cells

et al. 2018

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Purpose Prolonged bed rest and microgravity in space cause intervertebral disc (IVD) degeneration. However, the underlying molecular mechanisms are not completely understood. Transient receptor potential canonical (TRPC) channels are implicated in mechanosensing of several tissues, but are poorly explored in IVDs. Methods Primary human IVD cells from surgical biopsies composed of both annulus fibrosus and nucleus pulposus (passage 1-2) were exposed to simulated microgravity and to the TRPC channel inhibitor SKF-96365 (SKF) for up to 5 days. Proliferative capacity, cell cycle distribution, senescence and TRPC channel expression were analyzed. Results Both simulated microgravity and TRPC channel antagonism reduced the proliferative capacity of IVD cells and induced senescence. While significant changes in cell cycle distributions (reduction in G1 and accumulation in G2/M) were observed upon SKF treatment, the effect was small upon 3 days of simulated microgravity. Finally, downregulation of TRPC6 was shown under simulated microgravity. Conclusions Simulated microgravity and TRPC channel inhibition both led to reduced proliferation and increased senescence. Furthermore, simulated microgravity reduced TRPC6 expression. IVD cell senescence and mechanotransduction may hence potentially be regulated by TRPC6 expression. This study thus reveals promising targets for future studies.Graphical abstract These slides can be retrieved under Electronic Supplementary Material.

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A Novel Microgravity Simulator Applicable for Three-Dimensional Cell Culturing

Cited by 30 publications

References 29 publications

Validation of Random Positioning Versus Clinorotation Using a Macrophage Model System

Validation of Random Positioning Versus Clinorotation Using a Macrophage Model System

Simulated Microgravity: Critical Review on the Use of Random Positioning Machines for Mammalian Cell Culture

TRPC6 in simulated microgravity of intervertebral disc cells

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