Figure 2.

Preparation of muscle-on-a-chip devices for microgravity studies and experimental overview

(A) The overview of muscle-on-a-chip preparation consists of seeding human muscle cells onto immobilized aligned nanofibrillar collagen scaffolds followed by cell seeding. After one day, the cell-seeded scaffolds are placed into bioreactors and filled with growth media. The bioreactor is then secured with screws for transportation to the ISSNL.

(B) The experimental overview consists of incubating the bioreactor during transit to the ISSNL in plate habitats that provide temperature maintenance during transport to the ISSNL or in normal gravity conditions on Earth. Upon docking at the ISSNL, microgravity experiments began (day 0) with media change into an induction media that supports myotube formation. In some experiments, drugs were added to the media on day 0 and again on day 3. On day 7, the conditioned media were extracted, and the samples were stabilized in RNAlater or fixed in paraformaldehyde for analysis on by RNA sequencing, proteomics analysis, or myotube formation.