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Serum‐reduced media impacts on cell viability and protein expression in human lung epithelial cells
Serum starvation is a widely used condition in molecular biology experiments. Opti‐MEM is a serum‐reduced media used during transfection of genetic molecules into mammalian cells. However, the impact of such media on cell viability and protein synthesis is unknown. A549 human lung epithelial cell vi...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6519280/ https://www.ncbi.nlm.nih.gov/pubmed/30515823 http://dx.doi.org/10.1002/jcp.27890 |
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author | Rashid, Mahamud‐ur Coombs, Kevin M. |
author_facet | Rashid, Mahamud‐ur Coombs, Kevin M. |
author_sort | Rashid, Mahamud‐ur |
collection | PubMed |
description | Serum starvation is a widely used condition in molecular biology experiments. Opti‐MEM is a serum‐reduced media used during transfection of genetic molecules into mammalian cells. However, the impact of such media on cell viability and protein synthesis is unknown. A549 human lung epithelial cell viability and morphology were adversely affected by growing in Opti‐MEM. The cellular protein levels of chloride intracellular channel protein 1, proteasome subunit alpha Type 2, and heat shock 70 kDa protein 5 were dysregulated in A549 cells after growing in serum‐reduced media. Small interfering RNA transfection was done in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum, and knockdown efficacy was determined compared with Opti‐MEM. Similar amounts of knockdown of the target proteins were achieved in DMEM, and cell viability was higher compared with Opti‐MEM after transfection. Careful consideration of the impact of Opti‐MEM media during the culture or transfection is important for experimental design and results interpretation. |
format | Online Article Text |
id | pubmed-6519280 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-65192802019-05-23 Serum‐reduced media impacts on cell viability and protein expression in human lung epithelial cells Rashid, Mahamud‐ur Coombs, Kevin M. J Cell Physiol Rapid Communications Serum starvation is a widely used condition in molecular biology experiments. Opti‐MEM is a serum‐reduced media used during transfection of genetic molecules into mammalian cells. However, the impact of such media on cell viability and protein synthesis is unknown. A549 human lung epithelial cell viability and morphology were adversely affected by growing in Opti‐MEM. The cellular protein levels of chloride intracellular channel protein 1, proteasome subunit alpha Type 2, and heat shock 70 kDa protein 5 were dysregulated in A549 cells after growing in serum‐reduced media. Small interfering RNA transfection was done in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum, and knockdown efficacy was determined compared with Opti‐MEM. Similar amounts of knockdown of the target proteins were achieved in DMEM, and cell viability was higher compared with Opti‐MEM after transfection. Careful consideration of the impact of Opti‐MEM media during the culture or transfection is important for experimental design and results interpretation. John Wiley and Sons Inc. 2018-12-04 2019-06 /pmc/articles/PMC6519280/ /pubmed/30515823 http://dx.doi.org/10.1002/jcp.27890 Text en © 2018 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
spellingShingle | Rapid Communications Rashid, Mahamud‐ur Coombs, Kevin M. Serum‐reduced media impacts on cell viability and protein expression in human lung epithelial cells |
title | Serum‐reduced media impacts on cell viability and protein expression in human lung epithelial cells |
title_full | Serum‐reduced media impacts on cell viability and protein expression in human lung epithelial cells |
title_fullStr | Serum‐reduced media impacts on cell viability and protein expression in human lung epithelial cells |
title_full_unstemmed | Serum‐reduced media impacts on cell viability and protein expression in human lung epithelial cells |
title_short | Serum‐reduced media impacts on cell viability and protein expression in human lung epithelial cells |
title_sort | serum‐reduced media impacts on cell viability and protein expression in human lung epithelial cells |
topic | Rapid Communications |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6519280/ https://www.ncbi.nlm.nih.gov/pubmed/30515823 http://dx.doi.org/10.1002/jcp.27890 |
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