A Tailored Lipid Supplement Restored Membrane Fatty Acid Composition and Ameliorates In Vitro Biological Features of Human Amniotic Epithelial Cells

Abstract

Cell culture conditions influence several biological and biochemical features of stem cells (SCs), including the membrane lipid profile, thus limiting the use of SCs for cell therapy approaches. The present study aims to investigate whether the in vitro culture may alter the membrane fatty acid signature of human Amniotic Epithelial Cells (hAECs). The analysis of the membrane fatty acid composition of hAECs cultured in basal medium showed a loss in polyunsaturated fatty acids (PUFA), in particular in omega-6 (ω-6) content, compared to freshly isolated hAECs. The addition to the basal culture medium of a chemically defined and animal-free tailored lipid supplement, namely Refeed®, partially restored the membrane fatty acid signature of hAECs. Although the amelioration of the membrane composition did not prolong hAECs culture lifespan, Refeed® influenced cell morphology, counteracted the onset of senescence, and increased the migratory capacity as well as the ability of hAECs to inhibit Peripheral Blood Mononuclear Cell (PBMC) proliferation. This study provides new information on hAEC features during culture passages and demonstrates that the maintenance of the membrane fatty acid signature preserved higher cell quality during in vitro expansion, suggesting the use of lipid supplementation for SC expansion in cell-based therapies.

Trappc9 Deficiency Impairs the Plasticity of Stem Cells

“…For an example, excessive ROS accumulated in trappc9-deficient ASCs might constantly damage DNA molecules and subsequently initiate cellular senescence [51]. A study reported that lipid balance was important to delaying the cellular senescence of human amniotic epithelial cells [52]. Hence, lipid metabolism disturbance seen in trappc9null ASCs may contribute significantly to the premature cellular senescence.…”

Plumbagin, a Natural Compound with Several Biological Effects and Anti-Inflammatory Properties

“…However, they need to be expanded in vitro to obtain a sufficient number of cells for transplantation. During the in vitro culturing, stem cells undergo replicative senescence [ 87 , 88 ], a process that can be triggered by elevated ROS levels [ 89 , 90 ]. In 2013, a protective role of PB against senescence in human amniotic stem cells was proposed [ 89 ].…”

Mitigating Oxidative Stress in Perinatal Cells: A Critical Step toward an Optimal Therapeutic Use in Regenerative Medicine

“…Regarding the relevance of the culture medium, we also reported that during in vitro culture in traditional medium, AECs lose their native membrane lipid profile, with a decrease in polyunsaturated fatty acid (PUFA) content and particularly in omega 6. The changes in the AEC fatty acid profile caused the onset of senescent features, including an increase in cell dimension and the expression of senescence-associated genes such as p16 [ 35 ].…”

“…The addition of a lipid supplement restored the AEC membrane fatty acid content, increasing the omega-6 percentage to physiological values. The recovery of an in vivo-like membrane fatty acid profile delayed the onset of senescence features and enhanced the immunomodulatory ability of AECs compared with standard culture conditions [ 35 ].…”