Heat-Inactivation of Fetal and Newborn Sera Did Not Impair the Expansion and Scaffold Engineering Potentials of Fibroblasts

Pellerin, Félix-Antoine; Caneparo, Christophe; Pellerin, Ève; Chabaud, Stéphane; Pelletier, Martin; Bolduc, Stéphane

doi:10.3390/bioengineering8110184

Cited by 5 publications

(6 citation statements)

References 35 publications

(40 reference statements)

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“…A point that should be clarified in the protocols of FBS preparation for in vitro EV studies regards the heat‐inactivation procedure. Many studies do not specify whether FBS was heat inactivated (usually 56°C for 30–60 min under agitation (Bruinink et al., 2004; Giard, 1987; Leshem et al., 1999; Pellerin et al., 2021; Soltis et al., 1979)) prior to use, and when specified the authors usually do not explain if the heat inactivation procedure was performed before or after the EV depletion or how the heat inactivation was performed in terms of temperature and time. To better standardize EV research, information concerning the use of FBS in cell cultures should be added in the methods section of published works.…”

Section: Fbs Contaminants In Cell‐derived Ev Samples and Possible Sol...mentioning

confidence: 99%

“…Generally, before being added to the cell medium FBS is heat inactivated at 56°C for 30–60 minutes to destroy complement activity and inactivate potential microbial contaminants (Soltis et al., 1979; Triglia & Linscott, 1980). The removal of complement activity from FBS is required for cell cultures that are sensitive to it, such as immune cells, while it is unnecessary for others (Fante et al., 2021; Pellerin et al., 2021). Heat inactivation does not improve the growth promotion ability of FBS and may have adverse effects; for instance, it can reduce the ability of FBS to promote the cell attachment (Giard, 1987), neurite growth (Bird & Owen, 1998), proliferation, and metabolism of mesenchymal stromal cells (Tonarova et al., 2021).…”

Section: Fbs: Pros and Cons For Cell Culturesmentioning

confidence: 99%

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The dark side of foetal bovine serum in extracellular vesicle studies

Urzì

Bagge

Crescitelli

2022

J of Extracellular Vesicle

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Extracellular vesicles (EVs) have been shown to be involved in cell‐cell communication and to take part in both physiological and pathological processes. Thanks to their exclusive cargo, which includes proteins, lipids, and nucleic acids from the originating cells, they are gaining interest as potential biomarkers of disease. In recent years, their appealing features have been fascinating researchers from all over the world, thus increasing the number of in vitro studies focused on EV release, content, and biological activities. Cultured cell lines are the most‐used source of EVs; however, the EVs released in cell cultures are influenced by the cell culture conditions, such as the use of foetal bovine serum (FBS). FBS is the most common supplement for cell culture media, but it is also a source of contaminants, such as exogenous bovine EVs, RNA, and protein aggregates, that can contaminate the cell‐derived EVs and influence their cargo composition. The presence of FBS contaminants in cell‐derived EV samples is a well‐known issue that limits the clinical applications of EVs, thus increasing the need for standardization. In this review, we will discuss the pros and cons of using FBS in cell cultures as a source of EVs, as well as the protocols used to remove contaminants from FBS.

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Section: Fbs Contaminants In Cell‐derived Ev Samples and Possible Sol...mentioning

confidence: 99%

Section: Fbs: Pros and Cons For Cell Culturesmentioning

confidence: 99%

The dark side of foetal bovine serum in extracellular vesicle studies

Urzì

Bagge

Crescitelli

2022

J of Extracellular Vesicle

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“…The medium, supplemented or not with BPs, was changed daily for three days. On days 1 to 3, cells from three wells were collected with trypsin, centrifuged at 300 g for 10 min, resuspended in 10 mL ISOTON II diluent (Beckman Coulter, Mississauga, ON, Canada) and counted separately using a Z2 Coulter Particle Count and Size Analyzer (Beckman Coulter) [ 26 ]. A graph illustrating the numbers of cells per well as a function of time was performed to calculate the proliferation rate.…”

Section: Methodsmentioning

confidence: 99%

Bisphenols A and S Alter the Bioenergetics and Behaviours of Normal Urothelial and Bladder Cancer Cells

Pellerin

Chabaud

et al. 2022

Cancers

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Bisphenol A (BPA) and bisphenol S (BPS) are used in the production of plastics. These endocrine disruptors can be released into the environment and food, resulting in the continuous exposure of humans to bisphenols (BPs). The bladder urothelium is chronically exposed to BPA and BPS due to their presence in human urine samples. BPA and BPS exposure has been linked to cancer progression, especially for hormone-dependent cancers. However, the bladder is not recognized as a hormone-dependent tissue. Still, the presence of hormone receptors on the urothelium and their role in bladder cancer initiation and progression suggest that BPs could impact bladder cancer development. The effects of chronic exposure to BPA and BPS for 72 h on the bioenergetics (glycolysis and mitochondrial respiration), proliferation and migration of normal urothelial cells and non-invasive and invasive bladder cancer cells were evaluated. The results demonstrate that chronic exposure to BPs decreased urothelial cells’ energy metabolism and properties while increasing them for bladder cancer cells. These findings suggest that exposure to BPA and BPS could promote bladder cancer development with a potential clinical impact on bladder cancer progression. Further studies using 3D models would help to understand the clinical consequences of this exposure.

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“…The protocol to determine cellular metabolism has already been described [ 45 , 46 ]. Briefly, fibroblasts and epithelial cells (200,000 cells/cm 2 in 100 μL cell culture medium/well) were seeded in XFe96 96-well plates.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of a Serum-Free Medium for Human Epithelial and Stromal Cell Culture

Caneparo

Chabaud

Fradette

et al. 2022

IJMS

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Over the past decade, growing demand from many domains (research, cosmetics, pharmaceutical industries, etc.) has given rise to significant expansion of the number of in vitro cell cultures. Despite the widespread use of fetal bovine serum, many issues remain. Among them, the whole constitution of most serums remains unknown and is subject to significant variations. Furthermore, the presence of potential contamination and xenogeny elements is challenging for clinical applications, while limited production is an obstacle to the growing demand. To circumvent these issues, a Serum-Free Medium (SFM) has been developed to culture dermal and vesical fibroblasts and their corresponding epithelial cells, namely, keratinocytes and urothelial cells. To assess the impact of SFM on these cells, proliferation, clonogenic and metabolic assays have been compared over three passages to conditions associated with the use of a classic Fetal Bovine Serum-Containing Medium (FBSCM). The results showed that the SFM enabled fibroblast and epithelial cell proliferation while maintaining a morphology, cell size and metabolism similar to those of FBSCM. SFM has repeatedly been found to be better suited for epithelial cell proliferation and clonogenicity. Fibroblasts and epithelial cells also showed more significant mitochondrial metabolism in the SFM compared to the FBSCM condition. However, the SFM may need further optimization to improve fibroblast proliferation.

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Heat-Inactivation of Fetal and Newborn Sera Did Not Impair the Expansion and Scaffold Engineering Potentials of Fibroblasts

Cited by 5 publications

References 35 publications

The dark side of foetal bovine serum in extracellular vesicle studies

The dark side of foetal bovine serum in extracellular vesicle studies

Bisphenols A and S Alter the Bioenergetics and Behaviours of Normal Urothelial and Bladder Cancer Cells

Evaluation of a Serum-Free Medium for Human Epithelial and Stromal Cell Culture

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