Simulated microgravity modifications in musculoskeletal cells

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ICE Krakow

ICE Krakow

ul. Marii Konopnickiej 17 30-302 Kraków


Campioli, Alessio (DIMES, University of Genoa, Italy)


Gravity is one of the four fundamental forces that govern the universe and that we experience in daily life without realizing it.
The absence of mechano-stimulation caused by the lack or reduced presence of gravity (microgravity) has been widely studied since the beginning of the space age. This condition creates various cascade reactions in human, resulting in a profound remodeling and adaptation of organs and tissues in the organism. In particular, osteopenia and sarcopenia are two of the first changes observed after space flights that impact deeply astronaut wellness.

We are studying the effects of microgravity in vitro through the Random Positioning Machine (RPM), a tool capable of simulating it. This instrument has a plane that moves on two rotation axes at a random speed and direction leading to the nullification, averaged over time, of the gravity vector. The cells we are studying are C2C12 and MLO-Y4 which are respectively myoblast and murine osteocyte cell lines.

Methodology & results
In the C2C12 cell line we started to study the expression of differentiation genes such as MyoD, MyoG and the different isoforms of MHC at different times of culture (1, 3, 5 and 7 days) in ground control and on RPM. Through Real time PCR analysis of early and late differentiation marker genes it has been seen that this condition leads to a slowing of differentiation process which becomes statistically significant at 5 days in the case of MHC 2 alpha mRNA expression (P≤0.05).

In the case of the osteocyte cell line MLO-Y4, we noticed cell behavior changes after 5 days in RPM. In particular, connexin 43, a protein that forms gap junctions between cells with key functions in signal transduction and in response to hormonal and mechanical stimuli was downregulated in RPM (P ≤ 0.0001). Furthermore, genes linked to cellular senescence such as P53 (tumor suppressor), P21 and P16 were found to be much reduced in simulated microgravity (P53 = P ≤ 0.01, P21 = P ≤ 0.0001). Finally, the quantification of BAX mRNA, factor that regulates cell death, showed a reduced expression in RPM statistically significant (P ≤ 0.001).

Further experiments will be needed to fully understand the complex connection that links the expression of these genes and microgravity exposure.

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