title: Use of cyclodextrins for manipulating cellular cholesterol content.
abstract: 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.
authors.affiliation: Department of Biochemistry, Allegheny University of the Health Sciences, Philadelphia, PA 19129, USA.
authors.forename: A E
authors.initials: AE
authors.lastname: Christian
authors.forename: M P
authors.initials: MP
authors.lastname: Haynes
authors.forename: M C
authors.initials: MC
authors.lastname: Phillips
authors.forename: G H
authors.initials: GH
authors.lastname: Rothblat
mesh_terms: D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000818:Animals; D016466:CHO Cells; D006528:Carcinoma, Hepatocellular; D002784:Cholesterol / Q000378:metabolism*; D006224:Cricetinae; D003505:Cyclodextrins / Q000032:analysis / Q000494:pharmacology*; D051381:Rats; D012995:Solubility; D002785:Sterol O-Acyltransferase / Q000378:metabolism; D014407:Tumor Cells, Cultured; D047392:beta-Cyclodextrins
chemical_list: D003505:Cyclodextrins; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D000073738:2-Hydroxypropyl-beta-cyclodextrin; D002784:Cholesterol; D002785:Sterol O-Acyltransferase