Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Rashid, Mahamud‐ur, Coombs, Kevin M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
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
_version_ 1783418613519089664
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
work_keys_str_mv AT rashidmahamudur serumreducedmediaimpactsoncellviabilityandproteinexpressioninhumanlungepithelialcells
AT coombskevinm serumreducedmediaimpactsoncellviabilityandproteinexpressioninhumanlungepithelialcells