Evaluation of Simulated Microgravity Environments Induced by Diamagnetic Levitation of Plant Cell Suspension Cultures
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Ground-Based Facilities (GBF) are essetial tools to understand the physical and biological effects of the absence of gravity and they are necessary to prepare and complement space experiments. It has been shown previously that a real microgravity environment induces the dissociation of cell proliferation from cell growth in seedling root meristems, which are limited populations of proliferating cells. Plant cell cultures are large and homogeneous populations of proliferating cells, so that they are a convenient model to study the effects of altered gravity on cellular mechanisms regulating cell proliferation and associated cell growth. Cell suspension cultures of the Arabidopsis thaliana cell line MM2d were exposed to four altered gravity and magnetic field environments in a magnetic levitation facility for 3 hours, including two simulated microgravity and Mars-like gravity levels obtained with different magnetic field intensities. Samples were processed either by quick freezing, to be used in flow cytometry for cell cycle studies, or by chemical fixation for microscopy techniques to measure parameters of the nucleolus. Although the trend of the results was the same as those obtained in real microgravity on meristems (increased cell proliferation and decreased cell growth), we provide a technical discussion in the context of validation of proper conditions to achieve true cell levitation inside a levitating droplet. We conclude that the use of magnetic levitation as a simulated microgravity GBF for cell suspension cultures is not recommended.
KeywordsSimulated microgravity Suspension cell culture Magnetic levitation Ground-based facilities Arabidopsis thaliana Cell growth Cell proliferation Nucleolus
We wish to thank Dr. Julio Sáez-Vásquez (CNRS-University of Perpignan-Via Domitia, Perpignan, France) for his generous supply of anti-nucleolin antibody. This work was supported by grants of the Spanish National Plan for Research and Development, Ref. Nos. AYA2010-11834-E, and AYA2012-33982, access to Magnet facilities by the European Union (EUROMAGNET II) Project 2010.17 (NSO06-209) to FJM, the GBF project #4200022650 and #4000105761 to RH and ESA grant contract 4000107455112/NL/PA to JvL. KYK was supported by the Spanish CSIC JAE-PreDoc Program (Ref. JAEPre_2010_01894).
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