Document Detail


Introduction of phospholipids to cultured cells with cyclodextrin.
MedLine Citation:
PMID:  20881052     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Previous studies indicate that methyl-β-cyclodextrin (meβ-CD) can greatly enhance translocation of long-chain phospholipids from vesicles to cells in culture, which is very useful when studying, e.g., phospholipid metabolism and trafficking. However, the parameters affecting the transfer have not been systematically studied. Therefore, we studied the relevant parameters including meβ-CD and vesicle concentration, incubation time, phospholipid structure, and cell type. Because meβ-CD can extract cholesterol and other lipids from cells, thereby potentially altering cell growth or viability, these issues were studied as well. The results show that efficient incorporation of phospholipid species with hydrophobicity similar to that of natural species can be obtained without significantly compromising cell growth or viability. Cellular content of phosphatidyl-serine, -ethanolamine, and -choline could be increased dramatically, i.e., 400, 125, and 25%, respectively. Depletion of cellular cholesterol could be prevented or alleviated by inclusion of the proper amount of cholesterol in the donor vesicles. In summary, meβ-CD mediates efficient transfer of long-chain (phospho) lipids from vesicles to cells without significantly compromising their growth or viability. This lays a basis for detailed studies of phospholipid metabolism and trafficking as well as enables extensive manipulation of cellular phospholipid composition, which is particularly useful when investigating mechanisms underlying phospholipid homeostasis.
Authors:
Ville Kainu; Martin Hermansson; Pentti Somerharju
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-09-29
Journal Detail:
Title:  Journal of lipid research     Volume:  51     ISSN:  0022-2275     ISO Abbreviation:  J. Lipid Res.     Publication Date:  2010 Dec 
Date Detail:
Created Date:  2010-11-10     Completed Date:  2011-02-22     Revised Date:  2013-07-03    
Medline Journal Info:
Nlm Unique ID:  0376606     Medline TA:  J Lipid Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3533-41     Citation Subset:  IM    
Affiliation:
Institute of Biomedicine, Department of Medical Biochemistry and Developmental Biology, University of Helsinki, Helsinki, Finland. juho.kainu@helsinki.fi
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Proliferation / drug effects*
Cell Survival / drug effects
Cells, Cultured
Cholesterol / metabolism
Cricetinae
Cyclodextrins / pharmacology*
Fibroblasts / cytology,  metabolism
HeLa Cells / cytology,  metabolism
Homeostasis / physiology
Humans
Hydrophobic and Hydrophilic Interactions
Kidney / cytology,  embryology,  metabolism
Liposomes / metabolism*
Phospholipids / metabolism*
Spectrometry, Mass, Electrospray Ionization
Time Factors
beta-Cyclodextrins / pharmacology
Chemical
Reg. No./Substance:
0/Cyclodextrins; 0/Liposomes; 0/Phospholipids; 0/beta-Cyclodextrins; 0/methyl-beta-cyclodextrin; 57-88-5/Cholesterol
Comments/Corrections

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