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The amount of membrane cholesterol required for robust cell adhesion and proliferation in serum-free condition
Serum-containing medium is widely used to support cell attachment, stable growth and serial passaging of various cancer cell lines. However, the presence of cholesterols and lipids in serum greatly hinders the analysis of the effects of cholesterol depletion on cells in culture. In this study, we de...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9299302/ https://www.ncbi.nlm.nih.gov/pubmed/35857759 http://dx.doi.org/10.1371/journal.pone.0259482 |
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author | Takii, Shino Wu, Jun Okamura, Daiji |
author_facet | Takii, Shino Wu, Jun Okamura, Daiji |
author_sort | Takii, Shino |
collection | PubMed |
description | Serum-containing medium is widely used to support cell attachment, stable growth and serial passaging of various cancer cell lines. However, the presence of cholesterols and lipids in serum greatly hinders the analysis of the effects of cholesterol depletion on cells in culture. In this study, we developed a defined serum-free culture condition accessible to a variety of different types of adherent cancer cells. We tested different factors that are considered essential for cell culture and various extracellular matrix for plate coating, and found cells cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) basal media supplemented with Albumin (BSA) and insulin-transferrin-selenium-ethanolamine (ITS-X) on fibronectin-precoated plate (called as “DA-X condition”) showed comparable proliferation and survival to those in a serum-containing medium. Interestingly, we observed that DA-X condition could be adapted to a wide variety of adherent cancer cell lines, which enabled the analysis of how cholesterol depletion affected cancer cells in culture. Mechanistically, we found the beneficial effects of the DA-X condition in part can be attributed to the appropriate level of membrane cholesterol, and fibronectin-mediated signaling plays an important role in the suppression of cholesterol production. |
format | Online Article Text |
id | pubmed-9299302 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-92993022022-07-21 The amount of membrane cholesterol required for robust cell adhesion and proliferation in serum-free condition Takii, Shino Wu, Jun Okamura, Daiji PLoS One Research Article Serum-containing medium is widely used to support cell attachment, stable growth and serial passaging of various cancer cell lines. However, the presence of cholesterols and lipids in serum greatly hinders the analysis of the effects of cholesterol depletion on cells in culture. In this study, we developed a defined serum-free culture condition accessible to a variety of different types of adherent cancer cells. We tested different factors that are considered essential for cell culture and various extracellular matrix for plate coating, and found cells cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) basal media supplemented with Albumin (BSA) and insulin-transferrin-selenium-ethanolamine (ITS-X) on fibronectin-precoated plate (called as “DA-X condition”) showed comparable proliferation and survival to those in a serum-containing medium. Interestingly, we observed that DA-X condition could be adapted to a wide variety of adherent cancer cell lines, which enabled the analysis of how cholesterol depletion affected cancer cells in culture. Mechanistically, we found the beneficial effects of the DA-X condition in part can be attributed to the appropriate level of membrane cholesterol, and fibronectin-mediated signaling plays an important role in the suppression of cholesterol production. Public Library of Science 2022-07-20 /pmc/articles/PMC9299302/ /pubmed/35857759 http://dx.doi.org/10.1371/journal.pone.0259482 Text en © 2022 Takii et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Takii, Shino Wu, Jun Okamura, Daiji The amount of membrane cholesterol required for robust cell adhesion and proliferation in serum-free condition |
title | The amount of membrane cholesterol required for robust cell adhesion and proliferation in serum-free condition |
title_full | The amount of membrane cholesterol required for robust cell adhesion and proliferation in serum-free condition |
title_fullStr | The amount of membrane cholesterol required for robust cell adhesion and proliferation in serum-free condition |
title_full_unstemmed | The amount of membrane cholesterol required for robust cell adhesion and proliferation in serum-free condition |
title_short | The amount of membrane cholesterol required for robust cell adhesion and proliferation in serum-free condition |
title_sort | amount of membrane cholesterol required for robust cell adhesion and proliferation in serum-free condition |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9299302/ https://www.ncbi.nlm.nih.gov/pubmed/35857759 http://dx.doi.org/10.1371/journal.pone.0259482 |
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