abstract
This paper describes, for the first time to our knowledge, a lipidome and exometabolome characterization of osteogenic differentiation for human adipose tissue stem cells (hAMSCs) using nuclear magnetic resonance (NMR) spectroscopy. The holistic nature of NMR enabled the time-course evolution of cholesterol, mono- and polyunsaturated fatty acids (including omega-6 and omega-3 fatty acids), several phospholipids (phosphatidylcholine, phosphatidylethanolamine, sphingomyelins, and plasmalogens), and mono- and triglycerides to be followed. Lipid changes occurred almost exclusively between days 1 and 7, followed by a tendency for lipidome stabilization after day 7. On average, phospholipids and longer and more unsaturated fatty acids increased up to day 7, probably related to plasma membrane fluidity. Articulation of lipidome changes with previously reported polar endometabolome profiling and with exometabolome changes reported here in the same cells, enabled important correlations to be established during hAMSC osteogenic differentiation. Our results supported hypotheses related to the dynamics of membrane remodelling, anti-oxidative mechanisms, protein synthesis, and energy metabolism. Importantly, the observation of specific up-taken or excreted metabolites paves the way for the identification of potential osteoinductive metabolites useful for optimized osteogenic protocols.
keywords
NUCLEAR-MAGNETIC-RESONANCE; STROMAL CELLS; MASS-SPECTROMETRY; IN-VITRO; METABOLISM; NMR; ADIPOCYTES; H-1-NMR; ACID; OSTEOBLASTOGENESIS
subject category
Cell Biology
authors
Bispo, DSC; Michalkova, L; Correia, M; Jesus, CSH; Duarte, IF; Goodfellow, BJ; Oliveira, MB; Mano, JF; Gil, AM
our authors
Ana Maria Pissarra Coelho Gil
Associate Professor with Aggregation
Brian James Goodfellow
Assistant Professor
Daniela Bispo
PhD Student
Iola Melissa Fernandes Duarte
Principal Researcher
João Mano
Vice Director
Mariana Braga de Oliveira
Assistant ResearcherProjects
A Metabolomics-guided Bioreactor for Improved Engineered Bone Implants (BioImplant)
CICECO - Aveiro Institute of Materials (UIDB/50011/2020)
CICECO - Aveiro Institute of Materials (UIDP/50011/2020)
Associated Laboratory CICECO-Aveiro Institute of Materials (LA/P/0006/2020)
acknowledgements
The authors acknowledge the Portuguese Foundation for Science and Technology (FCT) for co-funding the BIOIMPLANT project (PTDC/BTM-ORG/28835/2017) through the COMPETE2020 program and European Union fund FEDER (POCI-01-0145-FEDER-028835). CSHJ is grateful to the same project for funding her contract with the University of Aveiro. DSB acknowledges the Sociedade Portuguesa de Quimica and FCT for her PhD grant SFRH/BD/150655/2020. This work was also financially supported by the European Research Council grant agreement ERC-2019-ADG-883370 (project REBORN). MBO acknowledges the individual contract CEECIND/03605/2017. This work was developed within the scope of the project CICECO-Aveiro Institute of Materials, UIDB/50011/2020, UIDP/50011/2020 & LA/P/0006/2020, financed by national funds through the FCT/MEC (PIDDAC). The NMR spectrometer used in this work is part of the National NMR Network (PTNMR) and, partially supported by Infrastructure Project N-022161 (co-financed by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC).