Effects of Simulated Microgravity on Wild Type and Marfan hiPSCs-Derived Embryoid Bodies

Abstract

Background Mechanical unloading in microgravity is thought to induce tissue degeneration by various mechanisms, including the inhibition of regenerative stem cell differentiation. In this work, we investigate the effects of microgravity simulation on early lineage commitment of hiPSCs from healthy and Marfan Syndrome (MFS; OMIM #154700) donors, using the embryoid bodies model of tissue differentiation and evaluating their ultra-structural conformation. MFS model involves an anomalous organization of the extracellular matrix for a deficit of fibrillin-1, an essential protein of connective tissue. Methods In vitro models require the use of embryoid bodies derived from hiPSCs. A DRPM was used to simulate microgravity conditions. Results Our data suggest an increase of the stemness of those EBs maintained in SMG condition. EBs are still capable of external migration, but are less likely to distinguish, providing a measure of the remaining progenitor or stem cell populations in the earlier stage. The microgravity response appears to vary between WT and Marfan EBs, presumably as a result of a cell structural component deficiency due to fibrillin-1 protein lack. In fact, MFS EBs show a reduced adaptive capacity to the environment of microgravity that prevented them from reacting and making rapid adjustments, while healthy EBs show stem retention, without any structural changes due to microgravity conditions. Conclusion EBs formation specifically mimics stem cell differentiation into embryonic tissues, this process has also significant similarities with adult stem cell-based tissue regeneration. The use of SMG devices for the maintenance of stem cells on regenerative medicine applications is becoming increasingly more feasible.

Single Cell in a Gravity Field

“…Similarly, after clinostating, spontaneous neuronal differentiation was observed to slow down: a quantitative shift of cells towards early neuroblasts (less differentiated cells) was noted compared to the control [ 35 ], the beating of 10-day-old embryoid bodies decreased against the background of changes in the expression of genes involved in heart morphogenesis, as well as encoding proteins of the MAP kinase cascade, focal adhesion, and cytoskeleton [ 37 ]. Embryoid bodies formed by induced human pluripotent stem cells hiPSC were also characterized by the maintenance of greater stemness in simulated microgravity [ 38 ]. At the same time, in contrast to spontaneous differentiation, activin A-directed differentiation of mESCs into the definitive endoderm was enhanced under conditions of simulated microgravity [ 39 ].…”

Unraveling the Relevance of Tissue‐Specific Decellularized Extracellular Matrix Hydrogels for Vocal Fold Regenerative Biomaterials: A Comprehensive Proteomic and In Vitro Study

“…Several of these proteins (EMILIN1, FBLN1, FBLN5, FBN1) contribute to elastic fiber formation, an essential process that repaired VF to regain elasticity. [33][34][35][36] Laminins, involved in basement membrane formation, integrin binding, and cell signaling were identified in all sample types (LAMA4, LAMB1, LAMB2, LAMC1), native VF only (LAMA3), or VF dECM only (LAMB3, LAMC2). [5,36] Seven proteoglycans were identified across all sample types.…”