Abstract
Embryonic developmental studies under microgravity conditions in space are very limited. To study the effects of short-term altered gravity on embryonic development processes, we exposed mouse embryonic stem cells (mESCs) to phases of hypergravity and microgravity and studied the differentiation potential of the cells using wide-genome microarray analysis. During the 64th European Space Agency's parabolic flight campaign, mESCs were exposed to 31 parabolas. Each parabola comprised phases lasting 22 s of hypergravity, microgravity, and a repeat of hypergravity. On different parabolas, RNA was isolated for microarray analysis. After exposure to 31 parabolas, mESCs (P31 mESCs) were further differentiated under normal gravity (1 g) conditions for 12 days, producing P31 12-day embryoid bodies (EBs). After analysis of the microarrays, the differentially expressed genes were analyzed using different bioinformatic tools to identify developmental and nondevelopmental biological processes affected by conditions on the parabolic flight experiment. Our results demonstrated that several genes belonging to GOs associated with cell cycle and proliferation were downregulated in undifferentiated mESCs exposed to gravity changes. However, several genes belonging to developmental processes, such as vasculature development, kidney development, skin development, and to the TGF-β signaling pathway, were upregulated. Interestingly, similar enriched and suppressed GOs were obtained in P31 12-day EBs compared with ground control 12-day EBs. Our results show that undifferentiated mESCs exposed to alternate hypergravity and microgravity phases expressed several genes associated with developmental/differentiation and cell cycle processes, suggesting a transition from the undifferentiated pluripotent to a more differentiated stage of mESCs.
Highlights
Increasing attention is being given on the influence of gravity on physiology and developmental biology
Given that nothing is known about the mechanical stress induced by alternate hypergravity and microgravity on the differentiation potential of mouse embryonic stem cells (mESCs), we investigated the effects of repeated short-term gravity alterations on their differentiation, induced by parabolic flight experiments
A simulated microgravity approach [18] and microarrays, we identified Gene Ontology categories (GOs) and KEGG pathways in differentiated mESCs affected by microgravity exposure [17]
Summary
Increasing attention is being given on the influence of gravity on physiology and developmental biology. In contrast to the clinostat, parabolic flight experiments facilitate investigation of the effects of acute (short period) and repeated exposure to hypergravity and microgravity on different cellular biological processes. We developed a so-called ‘‘pipette-based’’ methodology, suitable for mouse ESC (mESC) differentiation into embryoid bodies (EBs) within commercial plastic pipettes under 2D clinorotation [16] In this previous study, mESCs were differentiated using the pipette-method for 3 days under normal 1 g conditions, in contrast to simulated long-term microgravity induced by 3 days of clinorotation. Given that nothing is known about the mechanical stress induced by alternate hypergravity and microgravity on the differentiation potential of mESCs, we investigated the effects of repeated short-term gravity alterations on their differentiation, induced by parabolic flight experiments
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