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

Paola Spitalieri,Giuseppe Novelli,Maria Giovanna Scioli,Augusto Orlandi,Giuliana Longo,Mario Marini,Michela Murdocca,Federica Sangiuolo

doi:10.1007/s12195-021-00680-1

Paola Spitalieri, Giuseppe Novelli + Show 6 more

Open Access

https://doi.org/10.1007/s12195-021-00680-1

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Abstract

BackgroundMechanical 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.MethodsIn vitro models require the use of embryoid bodies derived from hiPSCs. A DRPM was used to simulate microgravity conditions.ResultsOur 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.ConclusionEBs 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.

Highlights

The capability of human induced Pluripotent Stem Cells to self-renew into undifferentiated cells and, at the same time, to differentiate into all cell types allows us to use them for studying cell differentiation process in a microgravity environment, in a way similar to what happens during fetal growth.[33]A key event called gastrulation occurs after implantation of the embryo into the uterus
We investigated the effects of microgravity simulation on early lineage commitment of human induced Pluripotent Stem Cells from healthy (WT) and Marfan Syndrome (MFS) donors, studying the embryoid bodies (EBs) model of tissue differentiation and their ultra-structural conformation
Morphological and Molecular Analyses in Embryoid Bodies (EBs) Cultured in Normal Gravity (1g) and Simulated Microgravity (SMG) Conditions

Summary

Introduction

A key event called gastrulation occurs after implantation of the embryo into the uterus During this stage of the mammalian development, the preimplantation embryo inner cell mass (ICM) constitutes three germ layers ectoderm (ecto = outer; such as skin and nerves) mesoderm (meso = middle; such as connective, bone and muscle), and endoderm (endo = inner; such as the gut tube and derivatives; like the lung and liver) and three body axes, head-tail, dorsal-ventral and left-right, are established. 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. Methods—In vitro models require the use of embryoid bodies derived from hiPSCs. A DRPM was used to simulate microgravity conditions.

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Conclusion