Clinorotation-induced weightlessness influences the cytoskeleton of glial cells in culture

Uva, Bianca Maria; Masini, Maria Angela; Sturla, Maddalena; Prato, Paola; Passalacqua, Mario; Giuliani, Massimo; Tagliafierro, Grazia; Strollo, Felice

doi:10.1016/s0006-8993(02)02415-0

Cited by 153 publications

(135 citation statements)

References 17 publications

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“…In fact, in FLG 29.1 cells exposed to the RPM, both Fas and FasL resulted overexpressed, compared to the controls, thus suggesting the triggering of the extrinsic pathway. In accordance with data reported by other authors [Grimm et al, 2002;Uva et al, 2002;Carlsson et al, 2003], we found cytoskeletal alterations as well. In particular, the cytoskeletal intermediate filament protein vimentin increased, while a-actin, F-actin, and tubulin decreased.…”

Section: Discussionsupporting

confidence: 93%

Modeled gravitational unloading triggers differentiation and apoptosis in preosteoclastic cells

et al. 2006

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Gravity acts permanently on organisms as either static or dynamic stimulation. Understanding the influence of gravitational and mechanical stimuli on biological systems is an intriguing scientific problem. More than two decades of life science studies in low g, either real or modeled by clinostats, as well as experimentation with devices simulating different types of controlled mechanical stimuli, have shown that important biological functions are altered at the single cell level. Here, we show that the human leukemic line FLG 29.1, characterized as an osteoclastic precursor model, is directly sensitive to gravitational unloading, modeled by a random positioning machine (RPM). The phenotypic expression of cytoskeletal proteins, osteoclastic markers, and factors regulating apoptosis was investigated using histochemical and immunohistochemical methods, while the expression of the corresponding genes was analyzed using RT-PCR. A quantitative bone resorption assay was performed. Autofluorescence spectroscopy and imaging were applied to gain information on cell metabolism. The results show that modeled hypogravity may trigger both differentiation and apoptosis in FLG 29.1 cells. Indeed, when comparing RPM versus 1 Â g cultures, in the former we found cytoskeletal alterations and a marked increase in apoptosis, but the surviving cells showed an osteoclastic-like morphology, overexpression of osteoclastic markers and the ability to resorb bone. In particular, the overexpression of both RANK and its ligand RANKL, maintained even after return to 1 Â g conditions, is consistent with the firing of a differentiation process via a paracrine/autocrine mechanism. J. Cell. Biochem. 98: 65-80, 2006. ß 2005 Wiley-Liss, Inc.Key words: weightlessness; preosteoclastic differentiation; apoptosis; bone resorption; osteoporosis; gene expressionThe importance of gravity in modulating some biological processes, such as plant gravitropism and the adaptation of the vertebrate skeleton in relation to its load-bearing function, has long been known. Nevertheless, the role of gravity and other mechanical factors in biological processes is far from clear. A weightless environment is a powerful tool for understanding this role and for studying the related cellular and molecular mechanisms. Several investigations conducted in space laboratories and with instruments averaging the gravity vector, and thus simulating conditions of reduced gravity, have shown that isolated cells may change their behavior under altered gravitational conditions. Loss of cell activation [Cogoli et al., 1984], perturbation of signal transduction [Schmitt et al., 1996;Hashemi et al., 1999] and modification of genetic expression [Walther et al., 1998;Hammond et al., 1999] have been described. This suggests that some of the mechanisms at the roots of the systemic effects observed in weightlessness, such as depression of the immune system [Gmü nder and Cogoli, 1996]

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Section: Discussionsupporting

confidence: 93%

Modeled gravitational unloading triggers differentiation and apoptosis in preosteoclastic cells

et al. 2006

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“…Recently, the RPM is used as a simulator for microgravity in various studies investigating responses in animal cells (for example Grimm et al 2002;Uva et al 2002;Infanger et al 2006;Pardo et al 2005;Patel et al 2009;Meloni et al 2006). However, the mode of operation of the RPM that was used in these studies varies.…”

Section: Discussionmentioning

confidence: 99%

“…The random rotation in all directions results in a net force of zero. Simulation of weightlessness with the RPM was reported to result in a wide variety of cellular responses which have also been found partly under real microgravity conditions (Hoson et al 1997;Schwarzenberg et al 1998;Grimm et al 2002;Uva et al 2002;Infanger et al 2006;Ulbrich et al 2008;Versari et al 2007a, b).…”

Section: Introductionmentioning

confidence: 93%

Simulation of Microgravity by Magnetic Levitation and Random Positioning: Effect on Human A431 Cell Morphology

Moes

Gielen

Bleichrodt

et al. 2010

Microgravity Sci. Technol.

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Simulation of weightlessness is a desired replenishment for research in microgravity since access to space flights is limited. In real microgravity conditions, the human epidermoid cell line A431 exhibits specific changes in the actin cytoskeleton resulting ultimately in the rounding-up of cells. This rounding of A431 cells was studied in detail during exposure to Random Positioning Machine (RPM) rotation and magnetic levitation. Random rotation and magnetic levitation induced similar changes in the actin morphology of A431 cells that were also described in real microgravity. A transient process of cell rounding and renewed spreading was observed in time, illustrated by a changing actin cytoskeleton and variation in the presence of focal adhesions. However, side effects of both methods easily can lead to false linking of cellular responses to simulated microgravity. Therefore further characterization of both methods is required.

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“…Most cells studied so far adjust to exposure to microgravity by altering the expression patterns of ECM and cytoskeletal proteins at gene and protein levels. [60][61][62][63][64] Preliminary indications of specific proteins involved in this process came from a comparison of the spheroid formation of the two thyroid cancer cell lines FTC-133 and CGTH-W1. The study revealed that speed of formation and size of resulting spheroids is related to the expression of certain proteins that bind fibronectin.…”

Section: Known Mechanisms Involved In Spheroid Formationmentioning

confidence: 99%

Growing Tissues in Real and Simulated Microgravity: New Methods for Tissue Engineering

Grimm

Wehland

Pietsch

et al. 2014

Tissue Engineering Part B: Reviews

124

137

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Tissue engineering in simulated (s-) and real microgravity (r-mg) is currently a topic in Space medicine contributing to biomedical sciences and their applications on Earth. The principal aim of this review is to highlight the advances and accomplishments in the field of tissue engineering that could be achieved by culturing cells in Space or by devices created to simulate microgravity on Earth. Understanding the biology of three-dimensional (3D) multicellular structures is very important for a more complete appreciation of in vivo tissue function and advancing in vitro tissue engineering efforts. Various cells exposed to r-mg in Space or to s-mg created by a random positioning machine, a 2D-clinostat, or a rotating wall vessel bioreactor grew in the form of 3D tissues. Hence, these methods represent a new strategy for tissue engineering of a variety of tissues, such as regenerated cartilage, artificial vessel constructs, and other organ tissues as well as multicellular cancer spheroids. These aggregates are used to study molecular mechanisms involved in angiogenesis, cancer development, and biology and for pharmacological testing of, for example, chemotherapeutic drugs or inhibitors of neoangiogenesis. Moreover, they are useful for studying multicellular responses in toxicology and radiation biology, or for performing coculture experiments. The future will show whether these tissue-engineered constructs can be used for medical transplantations. Unveiling the mechanisms of microgravity-dependent molecular and cellular changes is an up-to-date requirement for improving Space medicine and developing new treatment strategies that can be translated to in vivo models while reducing the use of laboratory animals.

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Clinorotation-induced weightlessness influences the cytoskeleton of glial cells in culture

Cited by 153 publications

References 17 publications

Modeled gravitational unloading triggers differentiation and apoptosis in preosteoclastic cells

Modeled gravitational unloading triggers differentiation and apoptosis in preosteoclastic cells

Simulation of Microgravity by Magnetic Levitation and Random Positioning: Effect on Human A431 Cell Morphology

Growing Tissues in Real and Simulated Microgravity: New Methods for Tissue Engineering

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