Characterization of Human Keratinocyte Cell Lines for Barrier Studies

Moran, Mary C.; Pandya, Radha P.; Leffler, Kimberly A.; Yoshida, Takeshi; Beck, Lisa A.; Brewer, Matthew G.

doi:10.1016/j.xjidi.2021.100018

Cited by 21 publications

(34 citation statements)

References 26 publications

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“…We have previously shown the state of KC differentiation impacts susceptibility to infection with vaccinia virus (VV) ( Moran et al, 2021 ). Since Western blot and TEER findings suggest differences in the kinetics of differentiation in cells propagated in Defined or KGM2 compared to KSFM media, we evaluated whether susceptibility to viral infection differed.…”

Section: Resultsmentioning

confidence: 99%

“…Cell Cultures: N/TERT2G cells were provided by Ellen H. van den Bogaard and grown to 30% confluency as previously described ( Dickson et al, 2000 , Moran et al, 2021 , Smits et al, 2017 ). Cells were cultured in one of three media; KSFM (Gibco, #17005042), KGM2 (PromoCell, #C-20211), and Defined (Gibco, #10744019).…”

Section: Methodsmentioning

confidence: 99%

“…Cell Cultures: N/TERT2G cells were provided by Ellen H. van den Bogaard and grown to 30% confluency as previously described (Dickson et al, 2000, Moran et al, 2021, Smits et al, 2017.…”

Section: Methodsmentioning

confidence: 99%

“…While within a lab the same media is commonly used, supply chain disruptions that occurred throughout the COVID-19 pandemic required our lab and labs around the globe to identify substitutions for many standard protocols. We found the use of alternative KC culture media gave us highly variable results in the immortalized KC line, N/TERT2G (Dickson et al, 2000, Moran et al, 2021.…”

Section: Introductionmentioning

confidence: 97%

See 3 more Smart Citations

Supply Chain Disruptions during COVID-19 Pandemic Uncover Differences in Keratinocyte Culture Media

Moran¹,

Pope²,

Brewer³

et al. 2022

JID Innovations

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Cell Cultures: N/TERT2G cells were provided by Ellen H. van den Bogaard and grown to 30% confluency as previously described (Dickson et al, 2000, Moran et al, 2021, Smits et al, 2017.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

See 2 more Smart Citations

Supply Chain Disruptions during COVID-19 Pandemic Uncover Differences in Keratinocyte Culture Media

Moran¹,

Pope²,

Brewer³

et al. 2022

JID Innovations

Self Cite

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“…For this reason, newly identified xanthones, even if less represented in the fruit, such as mangostanin, could show a higher antioxidant activity than other well-known molecules, explaining at least in part the superior effect of the total extract. This study employing HaCaT immortalized keratinocytes that have known limits to fully resemble skin needs to be validated in the future in primary keratinocytes, in order to account for donor-to-donor variability and in more sophisticated three-dimensional skin models [56]. To evaluate the topical availability of mangostanin in cosmeceutical formulations, it will be necessary to understand its penetration through the skin barrier and the resistance of the stratum corneum.…”

Section: Discussionmentioning

confidence: 99%

Mangostanin, a Xanthone Derived from Garcinia mangostana Fruit, Exerts Protective and Reparative Effects on Oxidative Damage in Human Keratinocytes

et al. 2022

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The fruit of Garcinia mangostana (mangosteen) is known in ancient traditional Asian medicine for its antioxidant, anti-inflammatory, immunomodulatory and anticancer activities. These effects are mainly due to the action of polyphenols known as xanthones, which are contained in the pericarp of the fruit. In recent years, there has been a growing interest from pharmaceutical companies in formulating new topicals based on mangosteen full extracts to prevent skin aging. However, the molecules responsible for these effects and the mechanisms involved have not been investigated so far. Here, the arils and shells of Garcinia mangostana were extracted with chloroform and methanol, and the extracts were further purified to yield 12 xanthone derivatives. Their effects were evaluated using in vitro cultures of human epidermal keratinocytes. After confirming the absence of cytotoxicity, we evaluated the antioxidant potential of these compounds, identifying mangostanin as capable of both protecting and restoring oxidative damage induced by H2O2. We showed how mangostanin, by reducing the generation of intracellular reactive oxygen species (ROS), prevents the activation of AKT (protein kinase B), ERK (extracellular signal-regulated kinase), p53, and other cellular pathways underlying cell damage and apoptosis activation. In conclusion, our study is the first to demonstrate that mangostanin is effective in protecting the skin from the action of free radicals, thus preventing skin aging, confirming a potential toward its development in the nutraceutical and cosmeceutical fields.

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Limonene protects human skin keratinocytes against UVB‐induced photodamage and photoaging by activating the Nrf2‐dependent antioxidant defense system

Kumar

Vani

Wang

2022

Environmental Toxicology

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Long term exposure to solar ultraviolet B (UVB) radiation is one of the primary factors of premature skin aging and is referred to as photoaging. Also, mammalian skin exposed to UVB triggers an increase in production of α-melanocyte-stimulating hormone (α-MSH), which is critically involved in the pathogenesis of hyperpigmentary skin diseases. This study investigated the protective effect of limonene on UVBinduced photodamage and photoaging in immortalized human skin keratinocytes (HaCaT) in vitro. Initially, we determined cell viability and levels of reactive oxygen species (ROS) in UVB-irradiated HaCaT cells. Pretreatment with limonene increased cell viability followed by inhibition of intracellular ROS generation in UVB-irradiated HaCaT cells. Interestingly, the antioxidative activity of limonene was directly correlated with an increase in expression of endogenous antioxidants, including heme oxygenase 1 (HO-1), NAD(P)H:quinone oxidoreductase 1 (NQO-1), and γ-glutamylcysteine synthetase (γ-GCLC), which was associated with enhanced nuclear translocation and activation of NF-E2-related factor-2 (Nrf2). Indeed, Nrf2 knockdown reduced limonene's protective effects. Additionally, we observed that limonene treatment inhibited UVB-induced α-MSH secretion followed by inhibition of proopiomelanocortin (POMC) via suppression of p53 transcriptional activation.Moreover, limonene prevented UVB-mediated depletion of tight junction regulatory proteins, including occludin and zonula occludens-1. On the other hand, limonene treatment significantly decreased matrix metalloproteinase-2 levels in UVB-irradiated HaCaT cells. Based on these results, limonene may have a dermato-protective effect in skin cells by activating the Nrf2-dependent cellular antioxidant defense system.

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Characterization of Human Keratinocyte Cell Lines for Barrier Studies

Cited by 21 publications

References 26 publications

Supply Chain Disruptions during COVID-19 Pandemic Uncover Differences in Keratinocyte Culture Media

Supply Chain Disruptions during COVID-19 Pandemic Uncover Differences in Keratinocyte Culture Media

Mangostanin, a Xanthone Derived from Garcinia mangostana Fruit, Exerts Protective and Reparative Effects on Oxidative Damage in Human Keratinocytes

Limonene protects human skin keratinocytes against UVB‐induced photodamage and photoaging by activating the Nrf2‐dependent antioxidant defense system

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