Abstract
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.
Highlights
Multicellular tumor spheroids (MCTS) formed scaffold-free under microgravity are of high interest for research and medicine
In order to prove ground-based microgravity simulation approaches, we investigated human follicular thyroid cancer cells (FTC-133) cultivated either on the CN or the random positioning machine (RPM) in a parallel manner focusing on the formation of spheroids as well as on alterations of gene expression and protein secretion
Spheroid formation on the RPM and the CN Subconfluent monolayers of human follicular thyroid carcinoma cells (FTC-133) were cultivated either on the RPM or on the CN and in parallel to the 1 g controls located in the same incubator, respectively
Summary
Multicellular 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. In. In order to understand and explain the effects of altered gravity on spheroid formation, we complemented our studies using two different ground-based facilities in order to simulate microgravity conditions – the 2D clinostat (CN) and the RPM. In order to understand and explain the effects of altered gravity on spheroid formation, we complemented our studies using two different ground-based facilities in order to simulate microgravity conditions – the 2D clinostat (CN) and the RPM Both devices are cost-efficient and enable a sufficient number of experiments, which is rarely achieved under real microgravity conditions [6]. The RPM consists of two independently rotating frames enabling a 3D rotation with random speed and random direction of the samples aiming to alter the influence of the gravity vector [8,9]
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