Cholesterol and phospholipid efflux from cultured cells.
The removal of phospholipids and cholesterol from tissues is the major mechanism mediating the initial assembly of high density lipoproteins (HDL), as well as being the main reason HDL are thought to protect against atherosclerosis. Investigations of the mechanisms of HDL assembly and testing of novel HDL-raising agents typically involve assays to determine phospholipid and/or cholesterol removal or "efflux" from cultured cells. The purpose of this chapter is to describe experimental protocols that can be used in the determination of cholesterol and phospholipid efflux from cultured cells by HDL apolipoproteins for the formation of new HDL particles, and the testing of novel HDL-raising therapies in vitro. A protocol is also provided for determining the size and nature of HDL particles formed in cell-conditioned medium using two-dimensional gel electrophoresis.
Waddington
Emma I
EI
CIHR Group on Molecular and Cell Biology of Lipids, Departments of Medicine and Biochemistry, University of Alberta, Edmonton, Alta., Canada T6G 2S2.
Boadu
Emmanuel
E
Francis
Gordon A
GA
D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D001161:Arteriosclerosis; D001671:Biochemistry / Q000379:methods*; D002460:Cell Line; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism*; D003470:Culture Media / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004587:Electrophoresis, Agar Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008075:Lipoproteins, HDL / Q000737:chemistry; D049294:Phospholipid Transfer Proteins / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism*
D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters; D003470:Culture Media; D017077:Culture Media, Conditioned; D008075:Lipoproteins, HDL; D049294:Phospholipid Transfer Proteins; D010743:Phospholipids; D002784:Cholesterol