Introduction of phospholipids to cultured cells with cyclodextrin
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 be...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2010-12-01
|
| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520313651 |
| _version_ | 1818727940999872512 |
|---|---|
| author | Ville Kainu Martin Hermansson Pentti Somerharju |
| author_facet | Ville Kainu Martin Hermansson Pentti Somerharju |
| author_sort | Ville Kainu |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-17T22:22:05Z |
| format | Article |
| id | doaj.art-1e0de7984b80415c9630261033641adb |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-17T22:22:05Z |
| publishDate | 2010-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-1e0de7984b80415c9630261033641adb2022-12-21T21:30:27ZengElsevierJournal of Lipid Research0022-22752010-12-01511235333541Introduction of phospholipids to cultured cells with cyclodextrinVille Kainu0Martin Hermansson1Pentti Somerharju2To whom correspondence should be addressed. juho.kainu@helsinki.fi; Institute of Biomedicine, Department of Medical Biochemistry and Developmental Biology, University of Helsinki, Helsinki, FinlandInstitute of Biomedicine, Department of Medical Biochemistry and Developmental Biology, University of Helsinki, Helsinki, FinlandInstitute of Biomedicine, Department of Medical Biochemistry and Developmental Biology, University of Helsinki, Helsinki, FinlandPrevious 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.http://www.sciencedirect.com/science/article/pii/S0022227520313651membranemetabolismremodelinghomeostasistrafficheavy-isotope |
| spellingShingle | Ville Kainu Martin Hermansson Pentti Somerharju Introduction of phospholipids to cultured cells with cyclodextrin Journal of Lipid Research membrane metabolism remodeling homeostasis traffic heavy-isotope |
| title | Introduction of phospholipids to cultured cells with cyclodextrin |
| title_full | Introduction of phospholipids to cultured cells with cyclodextrin |
| title_fullStr | Introduction of phospholipids to cultured cells with cyclodextrin |
| title_full_unstemmed | Introduction of phospholipids to cultured cells with cyclodextrin |
| title_short | Introduction of phospholipids to cultured cells with cyclodextrin |
| title_sort | introduction of phospholipids to cultured cells with cyclodextrin |
| topic | membrane metabolism remodeling homeostasis traffic heavy-isotope |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520313651 |
| work_keys_str_mv | AT villekainu introductionofphospholipidstoculturedcellswithcyclodextrin AT martinhermansson introductionofphospholipidstoculturedcellswithcyclodextrin AT penttisomerharju introductionofphospholipidstoculturedcellswithcyclodextrin |