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      Lipofuscin, Lipofuscin-Like Pigments and Autofluorescence

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          Abstract

          A brief overview is here provided on lipofuscin and lipofuscin-like substances, with particular reference to their biological significance as well as to their cellular origin and pathophysiological role. Special emphasis is also placed on the mutual relationships between lipofuscin and lipofuscin-like lipopigments on one side, and cell autofluorescence on the other.

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          Intracellular autofluorescence: a biomarker for epithelial cancer stem cells.

          Irene Miranda-Lorenzo, Jorge Dorado, Enza Lonardo … (2014)
          Cancer stem cells (CSCs) are thought to drive tumor growth, metastasis and chemoresistance. Although surface markers such as CD133 and CD44 have been successfully used to isolate CSCs, their expression is not exclusively linked to the CSC phenotype and is prone to environmental alteration. We identified cells with an autofluorescent subcellular compartment that exclusively showed CSC features across different human tumor types. Primary tumor-derived autofluorescent cells did not overlap with side-population (SP) cells, were enriched in sphere culture and during chemotherapy, strongly expressed pluripotency-associated genes, were highly metastatic and showed long-term in vivo tumorigenicity, even at the single-cell level. Autofluorescence was due to riboflavin accumulation in membrane-bounded cytoplasmic structures bearing ATP-dependent ABCG2 transporters. In summary, we identified and characterized an intrinsic autofluorescent phenotype in CSCs of diverse epithelial cancers and used this marker to isolate and characterize these cells.
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            Two-Photon Microscopy for Non-Invasive, Quantitative Monitoring of Stem Cell Differentiation

            William L. Rice, David L. Kaplan, Irene Georgakoudi (2010)
            Background The engineering of functional tissues is a complex multi-stage process, the success of which depends on the careful control of culture conditions and ultimately tissue maturation. To enable the efficient optimization of tissue development protocols, techniques suitable for monitoring the effects of added stimuli and induced tissue changes are needed. Methodology/Principal Findings Here, we present the quantitative use of two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) as a noninvasive means to monitor the differentiation of human mesenchymal stem cells (hMSCs) using entirely endogenous sources of contrast. We demonstrate that the individual fluorescence contribution from the intrinsic cellular fluorophores NAD(P)H, flavoproteins and lipofuscin can be extracted from TPEF images and monitored dynamically from the same cell population over time. Using the redox ratio, calculated from the contributions of NAD(P)H and flavoproteins, we identify distinct patterns in the evolution of the metabolic activity of hMSCs maintained in either propagation, osteogenic or adipogenic differentiation media. The differentiation of these cells is mirrored by changes in cell morphology apparent in high resolution TPEF images and by the detection of collagen production via SHG imaging. Finally, we find dramatic increases in lipofuscin levels in hMSCs maintained at 20% oxygen vs. those in 5% oxygen, establishing the use of this chromophore as a potential biomarker for oxidative stress. Conclusions/Significance In this study we demonstrate that it is possible to monitor the metabolic activity, morphology, ECM production and oxidative stress of hMSCs in a non-invasive manner. This is accomplished using generally available multiphoton microscopy equipment and simple data analysis techniques, such that the method can widely adopted by laboratories with a diversity of comparable equipment. This method therefore represents a powerful tool, which enables researchers to monitor engineered tissues and optimize culture conditions in a near real time manner.
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              Lipofuscin-bound iron is a major intracellular source of oxidants: role in senescent cells.

              Annika Höhn, Tobias Jung, Stefanie Grimm … (2010)
              Aging is accompanied by an intracellular accumulation of lipofuscin, a hydrophobic yellow-brownish material that accumulates especially in the lysosomal compartment, where it can be neither degraded nor exocytosed from the cell. The intracellular effects of accumulating lipofuscin are still a subject of speculation. In addition to the demonstrated inhibition of the proteasome, it was proposed that lipofuscin is cytotoxic because of its ability to incorporate transition metals such as copper and iron, resulting in a redox-active surface, able to catalyze the Fenton reaction. This characteristic of lipofuscin may contribute to an increased level of radical formation and oxidatively modified cellular components such as proteins, lipids, and RNA/DNA, which has been shown to be extensive in aging cells. In this study for the first time the lipofuscin-mediated formation of oxidants and the role of iron in this process are directly shown in a model of senescent fibroblasts, as well as in vitro with artificial lipofuscin. We demonstrate that this oxidant production is independent of mitochondria and has cytotoxic effects. The ability of lipofuscin to produce oxidants is dependent on the amount of transition metals incorporated. Although the amount of oxidants formed by cellular lipofuscin turned out to be moderate, it is chronic and thus lipofuscin is able to catalyze its own formation. Copyright 2010 Elsevier Inc. All rights reserved.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Eur J Histochem
                Journal ID (iso-abbrev): Eur J Histochem
                Journal ID (publisher-id): EJH
                Title: European Journal of Histochemistry : EJH
                Publisher: PAGEPress Publications, Pavia, Italy
                ISSN (Print): 1121-760X
                ISSN (Electronic): 2038-8306
                Publication date (Electronic): 06 February 2015
                Publication date Collection: 03 February 2015
                Volume: 59
                Issue: 1
                Electronic Location Identifier: 2485
                Affiliations
                Faculty of Veterinary Medicine, University of Teramo , Italy
                Author notes
                DVM, Dipl. ECVP, Faculty of Veterinary Medicine, University of Teramo, piazza Aldo Moro 45, 64100 Teramo, Italy. +39.861.266933 - +39.861.266865. gdiguardo@ 123456unite.it
                Article
                DOI: 10.4081/ejh.2015.2485
                PMC ID: 4378218
                PubMed ID: 25820564
                SO-VID: 333bc85e-ae68-432d-b042-b903fe832e02
                Copyright © ©Copyright G. Di Guardo
                License:

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                Date received : 10 January 2015
                Date accepted : 27 January 2015
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 22, Pages: 2
                Categories
                Subject: Views and Comments

                ScienceOpen disciplines: Clinical chemistry
                Keywords: lipofuscin,lipofuscin-like pigments,ceroid,ceroid lipofuscinosis,autofluorescence,fluorescence microscopy
                Data availability:
                ScienceOpen disciplines: Clinical chemistry
                Keywords: lipofuscin, lipofuscin-like pigments, ceroid, ceroid lipofuscinosis, autofluorescence, fluorescence microscopy

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