Abstract
Gravity plays a role in modulating plant growth and development and its alteration induces changes in these processes. Microgravity research has recently been extended to the use of in vitro plant cell cultures which are considered as an ideal model system to study cell proliferation and growth. In general, among the ground-based facilities available for microgravity simulation, the 2D pipette clinostat had been previously considered a suitable facility to be used for unicellular biological models although studies using single plant cell cultures raised some concerns. The incompatibility comes from the standard requirement of shaking a suspension culture for assuring its viability and active proliferation status in the control samples. Moreover, a related issue applies to the use of the random positioning machine (RPM) for cell suspension experiments. Here, we demonstrate an alternative culture method based on the immobilization of the culture before the altered gravity treatment occurs, such that it behaves as a solid object. Our immobilization procedure preserved plant cell culture viability without compromising basic cell properties as viability, morphology, cell cycle phases distribution, or chromatin organization, when compared with a standard cell suspension under shaking as a control. This approach should allow the space biology community to improve the quantity and quality of plant cell results in future simulated microgravity experiments or spaceflight opportunities.
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Acknowledgments
We wish to thank Dr. Crisanto Gutierrez at CBM (UAM-CSIC) for his generous supply of MM2d cultures and Dr. Jan Vos for his support in developing this culture technique. This work was supported by grants of the Spanish National Plan for Research and Development, Ref. Nos.AYA2012-33982 and ESP2015-64323-R, the Dutch Space Research Organization (NWO-ALW-SRON) [Grant number MG-057] and the ESA-ELIPS Program [ESA SEGMGSPE_Ph1 Project, contract number 4200022650]. KYK was supported by the Spanish CSIC JAE-PreDoc Program (Ref. JAEPre_2010_01894).
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Kamal, K.Y., van Loon, J.J.W.A., Medina, F.J. et al. Embedding Arabidopsis Plant Cell Suspensions in Low-Melting Agarose Facilitates Altered Gravity Studies. Microgravity Sci. Technol. 29, 115–119 (2017). https://doi.org/10.1007/s12217-016-9531-8
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DOI: https://doi.org/10.1007/s12217-016-9531-8