Adaptation of a 2-D Clinostat for Simulated Microgravity Experiments with Adherent Cells

Eiermann, Peter; Kopp, Sascha; Hauslage, Jens; Hemmersbach, Ruth; Gerzer, R.; Ivanova, Krassimira

doi:10.1007/s12217-013-9341-1

Cited by 55 publications

(60 citation statements)

References 18 publications

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“…Cells located at further distance from the rotation axis are exposed to accelerations reaching up to 0.036 g at about ±9 mm. Significant differences in gene expression within these acceleration intervals have been shown by Eiermann et al [30]. Based on these findings, only cells within the central 6 mm (≤0.012 g ) of a slide were harvested and analyzed in the present study.…”

Section: Discussionmentioning

confidence: 85%

Spheroid formation of human thyroid cancer cells under simulated microgravity: a possible role of CTGF and CAV1

et al. 2014

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BackgroundMulticellular tumor spheroids (MCTS) formed scaffold-free under microgravity are of high interest for research and medicine. Their formation mechanism can be studied in space in real microgravity or on Earth using ground-based facilities (GBF), which simulate microgravity. On Earth, these experiments are more cost-efficient and easily performable. However, each GBF might exert device-specific and altered superimposingly gravity-dependent effects on the cells.ResultsFTC-133 human thyroid cancer cells were cultivated on a 2D clinostat (CN) and a random positioning machine (RPM) and compared with corresponding 1 g control cells. Harvested cell samples were investigated by microscopy, quantitative realtime-PCR and Multi-Analyte Profiling. Spheroid formation and growth occurred during 72 h of cultivation on both devices. Cytokine secretion and gene activation patterns frequently altered in different ways, when the cells were cultured either on the RPM or the CN. A decreased expression of CAV1 and CTGF in MCTS compared to adherent cells was observed after cultivation on both machines.ConclusionThe development of MCTS proceeds similarly on the RPM and the CN resembling the situation observed under real microgravity conditions, while no MCTS formation was observed at 1 g under identical experimental conditions. Simultaneously, changes in the regulation of CTGF and CAV1 appeared in a comparable manner on both machines. A relationship between these molecules and MCTS formation is discussed.

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Section: Discussionmentioning

confidence: 85%

Spheroid formation of human thyroid cancer cells under simulated microgravity: a possible role of CTGF and CAV1

et al. 2014

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“…Ground‐based facilities: (A) 2D Fast‐rotating clinostat ( FRC ) as described by Eiermann et al . and used by Warnke et al . ; (B) 3D Random Positioning Machine ( RPM ) developed by ADS (Airbus, Defense & Space, former Dutch Space, Leiden, NL ) and described by van Loon ; (C) Rotating Wall Vessel ( RWV ) developed by NASA and described by Klaus et al .…”

Section: Ground‐based Facilities For Microgravity Simulation Used Formentioning

confidence: 99%

Scaffold‐free Tissue Formation Under Real and Simulated Microgravity Conditions

Aleshcheva

Bauer

Hemmersbach³

et al. 2016

Basic Clin Pharma Tox

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Scaffold-free tissue formation in microgravity is a new method in regenerative medicine and an important topic in Space Medicine. In this MiniReview, we focus on recent findings in the field of tissue engineering that were observed by exposing cells to real microgravity in space or to devices simulating to at least some extent microgravity conditions on Earth (ground-based facilities). Under both conditions -real and simulated microgravity -a part of the cultured cells of various populations detaches from the bottom of a culture flask. The cells form three-dimensional (3D) aggregates resembling the organs from which the cells have been derived. As spaceflights are rare and extremely expensive, cell culture under simulated microgravity allows more comprehensive and frequent studies on the scaffold-free 3D tissue formation in some aspects, as a number of publications have proven during the last two decades. In this MiniReview, we summarize data from our own studies and work from various researchers about tissue engineering of multi-cellular spheroids formed by cancer cells, tube formation by endothelial cells and cartilage formation by exposing the cells to ground-based facilities such as the 3D Random Positioning Machine (RPM), the 2D Fast-Rotating Clinostat (FRC) or the Rotating Wall Vessel (RWV). Subsequently, we investigated selforganization of 3D aggregates without scaffolds pursuing to enhance the frequency of 3D formation and to enlarge the size of the organ-like aggregates. The density of the monolayer exposed to real or simulated microgravity as well as the composition of the culture media revealed an impact on the results. Genomic and proteomic alterations were induced by simulated microgravity. Under microgravity conditions, adherent cells expressed other genes than cells grown in spheroids. In this MiniReview, the recent improvements in scaffold-free tissue formation are summarized and relationships between phenotypic and molecular appearance are highlighted.

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“…As the GC-cGMP pathway is involved in the melanocyte response to environmental stress and has also been linked to many cellular processes, including melanoma growth and migration, we compared the regulation of the GC-cGMP signaling pathway in a panel of human melanocytes and melanoma cell lines with different metastatic potential and pigmentation in hypergravity as well as under simulated microgravity conditions [80][81][82]. We used a specific centrifuge for the hypergravity experiments (up to 10 g) [80] and a fast-rotating 2D clinostat (60 rpm) as well as a fast-rotating clinostat microscope (60 rpm) to simulate microgravity conditions [81,83]. These devices were manufactured at the German Aerospace Center (DLR), Cologne, Germany.…”

Section: Differential Effects Of Altered Gravity On Guanylyl Cyclase-mentioning

confidence: 99%

Guanylyl Cyclase-cGMP Signaling Pathway in Melanocytes: Differential Effects of Altered Gravity in Non-Metastatic and Metastatic Cells

Ivanova

Hemmersbach

2020

IJMS

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Human epidermal melanocytes as melanin producing skin cells represent a crucial barrier against UV-radiation and oxidative stress. It was shown that the intracellular signaling molecule cyclic guanosine-3 ,5 -monophosphate (cGMP), generated by the guanylyl cyclases (GCs), e.g., the nitric oxide (NO)-sensitive soluble GC (sGC) and the natriuretic peptide-activated particulate GC (GC-A/GC-B), plays a role in the melanocyte response to environmental stress. Importantly, cGMP is involved in NO-induced perturbation of melanocyte-extracellular matrix interactions and in addition, increased NO production during inflammation may lead to loss of melanocytes and support melanoma metastasis. Further, the NO-sensitive sGC is expressed predominantly in human melanocytes and non-metastatic melanoma cells, whereas absence of functional sGC but up-regulated expression of GC-A/GC-B and inducible NO synthase (iNOS) are detected in metastatic cells. Thus, suppression of sGC expression as well as up-regulated expression of GC-A/GC-B/iNOS appears to correlate with tumor aggressiveness. As the cGMP pathway plays important roles in melanocyte (patho)physiology, we present an overview on the differential effects of altered gravity (hypergravity/simulated microgravity) on the cGMP signaling pathway in melanocytes and melanoma cells with different metastatic potential. We believe that future experiments in real microgravity may benefit from considering cGMP signaling as a possible factor for melanocyte transformation and in medication.

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Adaptation of a 2-D Clinostat for Simulated Microgravity Experiments with Adherent Cells

Cited by 55 publications

References 18 publications

Spheroid formation of human thyroid cancer cells under simulated microgravity: a possible role of CTGF and CAV1

Spheroid formation of human thyroid cancer cells under simulated microgravity: a possible role of CTGF and CAV1

Scaffold‐free Tissue Formation Under Real and Simulated Microgravity Conditions

Guanylyl Cyclase-cGMP Signaling Pathway in Melanocytes: Differential Effects of Altered Gravity in Non-Metastatic and Metastatic Cells

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