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.
Highlights
Lipid metabolism influences stem cell behavior by affecting cell proliferation and differentiation ability [1] (p. 115)
We further demonstrated how Refeed® supplementation affected hFM-mesenchymal/stromal stem cells (MSCs) paracrine features by impacting intracellular vesicle trafficking, exosome production, and secretive functions [21]. human Amniotic Epithelial Cells represent another cell population derived from placenta; in particular, hAECs derive from the amniotic membrane [17] and originate from pluripotent epiblast cells. hAECs express typical pluripotency markers, such as octamer-binding protein 4 (OCT4), SRY-related HMG-box gene 2 (SOX2), and Nanog [22]
Data show the percentage of BrdU incorporation of Peripheral Blood Mononuclear Cell (PBMC) co-cultured with hAECs with or without Refeed®, with respect to positive controls, PBMCs + PHA + Refeed® and PBMCs + PHA, respectively, both set at 100%
Summary
Lipid metabolism influences stem cell behavior by affecting cell proliferation and differentiation ability [1] (p. 115). The addition of Refeed® to the culture medium partially restored the membrane fatty acid signature and improved several characteristics of cultured hFM-MSCs, such as proliferative and immunomodulatory activity, as well as angiogenic differentiation capacity [9] The aim of this study was to investigate whether the in vitro culture may alter the membrane fatty acid signature of human Amniotic Epithelial Cells (hAECs). HAECs treated with Refeed® increased their migratory capacity, improved their ability to reduce the proliferation of Peripheral Blood Mononuclear Cells (PBMCs), and delayed the onset of senescence These data provided new information about hAEC features during cell culture and suggested that the optimization of culture conditions by the use of a tailored lipid supplement is able to preserve hAEC properties and improve the maintenance and the expansion process of cultured cells
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
