Restored in vivo-like membrane lipidomics positively influence in vitro features of cultured mesenchymal stromal/stem cells derived from human placenta.
The study of lipid metabolism in stem cell physiology has recently raised great interest. The role of lipids goes beyond the mere structural involvement in assembling extra- and intra-cellular compartments. Nevertheless, we are still far from understanding the impact of membrane lipidomics in stemness maintenance and differentiation patterns. In the last years, it has been reported how in vitro cell culturing can modify membrane lipidomics. The aim of the present work was to study the membrane fatty acid profile of mesenchymal stromal cells (MSCs) derived from human fetal membranes (hFM-MSCs) and to correlate this to specific biological properties by using chemically defined tailored lipid supplements (Refeed®).



Freshly isolated hFM-MSCs were characterized for their membrane fatty acid composition. hFM-MSCs were cultivated in vitro following a classical protocol and their membrane fatty acid profile at different passages was compared to the profile in vivo. A tailored Refeed® lipid supplement was developed with the aim of reducing the differences created by the in vitro cultivation and was tested on cultured hFM-MSCs. Cell morphology, viability, proliferation, angiogenic differentiation, and immunomodulatory properties after in vitro exposure to the tailored Refeed® lipid supplement were investigated.



A significant modification of hFM-MSC membrane fatty acid composition occurred during in vitro culture. Using a tailored lipid supplement, the fatty acid composition of cultured cells remained more similar to their in vivo counterparts, being characterized by a higher polyunsaturated and omega-6 fatty acid content. These changes in membrane composition had no effect on cell morphology and viability, but were linked with increased cell proliferation rate, angiogenic differentiation, and immunomodulatory properties. In particular, Refeed®-supplemented hFM-MSCs showed greater ability to express fully functional cell membrane molecules.



Culturing hFM-MSCs alters their fatty acid composition. A tailored lipid supplement is able to improve in vitro hFM-MSC functional properties by recreating a membrane environment more similar to the physiological counterpart. This approach should be considered in cell therapy applications in order to maintain a higher cell quality during in vitro passaging and to influence the outcome of cell-based therapeutic approaches when cells are administered to patients.
Remembrane Srl, Imola, Italy.
Alexandros
A
Chatgilialoglu
Department of Experimental, Diagnostic and Specialty Medicine, Unit of Histology, Embryology and Applied Biology, University of Bologna, Via Belmeloro 8, 40126, Bologna, Italy.
Martina
M
Rossi
Department of Experimental, Diagnostic and Specialty Medicine, Unit of Histology, Embryology and Applied Biology, University of Bologna, Via Belmeloro 8, 40126, Bologna, Italy. francesco.alviano@unibo.it.
Francesco
0000-0002-5315-0104
F
Alviano
Remembrane Srl, Imola, Italy.|Department of Morphology, Surgery and Experimental Medicine, Section of Pathology, Oncology and Experimental Biology, University of Ferrara, Ferrara, Italy.
Paola
P
Poggi
Department of Experimental, Diagnostic and Specialty Medicine, Unit of Histology, Embryology and Applied Biology, University of Bologna, Via Belmeloro 8, 40126, Bologna, Italy.|Department of Experimental, Diagnostic and Specialty Medicine, Unit of Nephrology, Dialysis and Renal Transplant, St, Orsola-Malpighi University Hospital, Via Massarenti 9, 40138, Bologna, Italy.
Chiara
C
Zannini
Department of Experimental, Diagnostic and Specialty Medicine, Unit of Histology, Embryology and Applied Biology, University of Bologna, Via Belmeloro 8, 40126, Bologna, Italy.
Cosetta
C
Marchionni
Service of Immunohematology and Transfusion Medicine, St. Orsola-Malpighi University Hospital, Via Massarenti 9, 40138, Bologna, Italy.
Francesca
F
Ricci
Service of Immunohematology and Transfusion Medicine, St. Orsola-Malpighi University Hospital, Via Massarenti 9, 40138, Bologna, Italy.
Pier Luigi
PL
Tazzari
Laboratory of Molecular Biology, Institute of Cardiology, Department of Experimental, Diagnostic and Specialty Medicine, St. Orsola - Malpighi University Hospital, University of Bologna, Via Massarenti 9, 40138, Bologna, Italy.|National Institute of Biostructures and Biosystems at Ettore Sansavini Health Science Foundation ONLUS - Lab SWITH, Corso Garibaldi 11, 48022, Lugo (RA), Italy.
Valentina
V
Taglioli
Human Development and Health Academic Unit, Faculty of Medicine, University of Southampton, Southampton, UK.|NIHR Southampton Biomedical Research Centre, University Hospital Southampton NHS Foundation Trust and University of Southampton, Southampton, UK.
Philip C
PC
Calder
Department of Experimental, Diagnostic and Specialty Medicine, Unit of Histology, Embryology and Applied Biology, University of Bologna, Via Belmeloro 8, 40126, Bologna, Italy.
Laura
L
Bonsi
D000975:Antioxidants / Q000494:pharmacology*; D002454:Cell Differentiation; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects*; D049109:Cell Proliferation; D019587:Dietary Supplements; D005321:Extraembryonic Membranes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005227:Fatty Acids / Q000032:analysis / Q000378:metabolism; D005229:Fatty Acids, Monounsaturated / Q000032:analysis / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000032:analysis / Q000378:metabolism; D005260:Female; D006801:Humans; D050356:Lipid Metabolism / Q000187:drug effects*; D008563:Membrane Lipids / Q000032:analysis / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010920:Placenta / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011247:Pregnancy; D061251:Primary Cell Culture
D000975:Antioxidants; D005227:Fatty Acids; D005229:Fatty Acids, Monounsaturated; D005231:Fatty Acids, Unsaturated; D008563:Membrane Lipids
Membrane fatty acids; Membrane lipidomics; Mesenchymal stromal cells; Stem cells