Cholesterol Efflux Assay

• DOI: 10.3791/3810
• Publication Date: 2012-03-06T18:50:55Z
• Authors: Hann Low; Anh Hoang; Dmitri Sviridov
• Keywords: Efflux

Cholesterol content of cells must be maintained within the very tight limits, too much or little cholesterol in a cell results disruption cellular membranes, apoptosis and necrosis 1. Cells can source from intracellular synthesis plasma lipoproteins, both sources are sufficient to fully satisfy cells' requirements for cholesterol. The processes uptake tightly regulated deficiencies rare 2. Excessive is more common problem 3. With exception hepatocytes some degree adrenocortical cells, unable degrade have two options reduce their content: convert into cholesteryl esters, an option with limited capacity as overloading esters also toxic, efflux, potentially unlimited capacity. efflux specific process that by number transporters, such ATP binding cassette transporter proteins A1 (ABCA1) G1 (ABCG1) scavenger receptor type B1. natural acceptor high density lipoprotein (HDL) apolipoprotein A-I. assay designed quantitate rate cultured cells. It measures maintain and/or acceptors accept released consists following steps. Step 1: labelling adding labelled serum-containing medium incubating 24-48 h. This step may combined loading 2: incubation serum-free equilibrate among all pools. stage activation transporters. 3: extracellular quantitation movement acceptor. If precursors were used label newly synthesized cholesterol, fourth step, purification required. delivers information: (i) how particular treatment (a mutation, knock-down, overexpression treatment) affects (ii) affected disease treatment. method often context cardiovascular research, metabolic neurodegenerative disorders, infectious reproductive diseases.