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Journal of Lipid Research, Vol 38, 2264-2272, Copyright © 1997 by Lipid Research, Inc.

ARTICLES

Use of cyclodextrins for manipulating cellular cholesterol content

AE Christian, MP Haynes, MC Phillips and GH Rothblat
Department of Biochemistry, Allegheny University of the Health Sciences, Philadelphia, PA 19129, USA.

Previous studies from this laboratory have demonstrated that exposure of tissue culture cells to cyclodextrins results in rapid cholesterol depletion. In the present study, we have developed experimental systems for using solutions of cyclodextrins, either 2-hydroxypropyl beta- cyclodextrin or methylated beta-cyclodextrin, complexed with varying amounts of free cholesterol to manipulate cell cholesterol content. Cholesterol delivered via the cyclodextrin has been found to be metabolically active, as measured by the acyl-coenzyme A:cholesterol acyltransferase (ACAT)-mediated esterification of [3H]cholesterol in Fu5AH rat hepatoma cells and Chinese hamster ovary cells. The methylated beta-cyclodextrin was found to be a more efficient donor in all cell types studied, with an average cholesterol uptake of at least 100 microg cholesterol/mg protein within 6 h. By modifying the cyclodextrin:cholesterol molar ratio, it is possible to manipulate the cellular cholesterol content of cells, producing conditions ranging from net cholesterol enrichment to depletion. The use of cyclodextrins provides a convenient, precise and reproducible method for modulating the cholesterol content of tissue culture cells.
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				A. M. Vaughan and J. F. Oram ABCA1 redistributes membrane cholesterol independent of apolipoprotein interactions J. Lipid Res., July 1, 2003; 44(7): 1373 - 1380. [Abstract] [Full Text] [PDF]

				S. M. Liu, A. Cogny, M. Kockx, R. T. Dean, K. Gaus, W. Jessup, and L. Kritharides Cyclodextrins differentially mobilize free and esterified cholesterol from primary human foam cell macrophages J. Lipid Res., June 1, 2003; 44(6): 1156 - 1166. [Abstract] [Full Text] [PDF]

				G. K. Hovingh, M. J. A. van Wijland, A. Brownlie, R. J. Bisoendial, M. R. Hayden, J. J. P. Kastelein, and A. K. Groen The role of the ABCA1 transporter and cholesterol efflux in familial hypoalphalipoproteinemia J. Lipid Res., June 1, 2003; 44(6): 1251 - 1255. [Abstract] [Full Text] [PDF]

				H. Wang, X. Jiang, F. Yang, J. W. Gaubatz, L. Ma, M. J. Magera, X. Yang, P. B. Berger, W. Durante, H. J. Pownall, et al. Hyperhomocysteinemia accelerates atherosclerosis in cystathionine beta -synthase and apolipoprotein E double knock-out mice with and without dietary perturbation Blood, May 15, 2003; 101(10): 3901 - 3907. [Abstract] [Full Text] [PDF]

				K. M. Wojtanik and L. Liscum The Transport of Low Density Lipoprotein-derived Cholesterol to the Plasma Membrane Is Defective in NPC1 Cells J. Biol. Chem., April 18, 2003; 278(17): 14850 - 14856. [Abstract] [Full Text] [PDF]

				H. Hering, C.-C. Lin, and M. Sheng Lipid Rafts in the Maintenance of Synapses, Dendritic Spines, and Surface AMPA Receptor Stability J. Neurosci., April 15, 2003; 23(8): 3262 - 3271. [Abstract] [Full Text] [PDF]

				B. Karten, D. E. Vance, R. B. Campenot, and J. E. Vance Trafficking of Cholesterol from Cell Bodies to Distal Axons in Niemann Pick C1-deficient Neurons J. Biol. Chem., January 31, 2003; 278(6): 4168 - 4175. [Abstract] [Full Text] [PDF]

				Y. Percherancier, B. Lagane, T. Planchenault, I. Staropoli, R. Altmeyer, J.-L. Virelizier, F. Arenzana-Seisdedos, D. C. Hoessli, and F. Bachelerie HIV-1 Entry into T-cells Is Not Dependent on CD4 and CCR5 Localization to Sphingolipid-enriched, Detergent-resistant, Raft Membrane Domains J. Biol. Chem., January 24, 2003; 278(5): 3153 - 3161. [Abstract] [Full Text] [PDF]

				K. Boesze-Battaglia, J. Dispoto, and M. A. Kahoe Association of a Photoreceptor-specific Tetraspanin Protein, ROM-1, with Triton X-100-resistant Membrane Rafts from Rod Outer Segment Disk Membranes J. Biol. Chem., October 25, 2002; 277(44): 41843 - 41849. [Abstract] [Full Text] [PDF]

				J. C. Cruz, M. Thomas, E. Wong, N. Ohgami, S. Sugii, T. Curphey, C. C. Y. Chang, and T.-Y. Chang Synthesis and biochemical properties of a new photoactivatable cholesterol analog 7,7-azocholestanol and its linoleate ester in Chinese hamster ovary cell lines J. Lipid Res., August 1, 2002; 43(8): 1341 - 1347. [Abstract] [Full Text] [PDF]

				C. Kunzelmann-Marche, J.-M. Freyssinet, and M. C. Martinez Loss of Plasma Membrane Phospholipid Asymmetry Requires Raft Integrity. ROLE OF TRANSIENT RECEPTOR POTENTIAL CHANNELS AND ERK PATHWAY J. Biol. Chem., May 24, 2002; 277(22): 19876 - 19881. [Abstract] [Full Text] [PDF]

				A. A. Wolf, Y. Fujinaga, and W. I. Lencer Uncoupling of the Cholera Toxin-GM1 Ganglioside Receptor Complex from Endocytosis, Retrograde Golgi Trafficking, and Downstream Signal Transduction by Depletion of Membrane Cholesterol J. Biol. Chem., May 3, 2002; 277(18): 16249 - 16256. [Abstract] [Full Text] [PDF]

				K. Gillotte-Taylor, M. Nickel, W. J. Johnson, O. L. Francone, P. Holvoet, S. Lund-Katz, G. H. Rothblat, and M. C. Phillips Effects of Enrichment of Fibroblasts with Unesterified Cholesterol on the Efflux of Cellular Lipids to Apolipoprotein A-I J. Biol. Chem., March 29, 2002; 277(14): 11811 - 11820. [Abstract] [Full Text] [PDF]

				A. Ahn, D. L. Gibbons, and M. Kielian The Fusion Peptide of Semliki Forest Virus Associates with Sterol-Rich Membrane Domains J. Virol., March 7, 2002; 76(7): 3267 - 3275. [Abstract] [Full Text] [PDF]

				S. Bavari, C. M. Bosio, E. Wiegand, G. Ruthel, A. B. Will, T. W. Geisbert, M. Hevey, C. Schmaljohn, A. Schmaljohn, and M. J. Aman Lipid Raft Microdomains: A Gateway for Compartmentalized Trafficking of Ebola and Marburg Viruses J. Exp. Med., March 4, 2002; 195(5): 593 - 602. [Abstract] [Full Text] [PDF]

				T. M. Holm, A. Braun, B. L. Trigatti, C. Brugnara, M. Sakamoto, M. Krieger, and N. C. Andrews Failure of red blood cell maturation in mice with defects in the high-density lipoprotein receptor SR-BI Blood, March 1, 2002; 99(5): 1817 - 1824. [Abstract] [Full Text] [PDF]

				C. Gajate and F. Mollinedo The antitumor ether lipid ET-18-OCH3 induces apoptosis through translocation and capping of Fas/CD95 into membrane rafts in human leukemic cells Blood, December 15, 2001; 98(13): 3860 - 3863. [Abstract] [Full Text] [PDF]

				J. Santiago, G. R. Guzman, L. V. Rojas, R. Marti, G. A. Asmar-Rovira, L. F. Santana, M. McNamee, and J. A. Lasalde-Dominicci Probing the Effects of Membrane Cholesterol in the Torpedo californica Acetylcholine Receptor and the Novel Lipid-exposed Mutation alpha C418W in Xenopus Oocytes J. Biol. Chem., November 30, 2001; 276(49): 46523 - 46532. [Abstract] [Full Text] [PDF]