Major cell biological parameters of keratinocytes are predetermined by culture medium and donor source

Zorn-Kruppa, Michaela; Volksdorf, Thomas; Ueck, Christopher; Zöller, Eva; Reinshagen, Konrad; Ridderbusch, Ina; Bruning, Guido; Houdek, Pia; Moll, Ingrid; Brandner, Johanna M.

doi:10.1111/exd.12922

Cited by 7 publications

(3 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cell culture. Primary human keratinocytes were isolated from foreskin as described before 31 Reconstructed human epidermis (RHE). 2 × 10 5 keratinocytes were seeded in 500 μL EpiLife medium (1.5 mM CaCl 2 ) onto Millicell cell culture inserts (0.4 μm, 12 mm diameter, Merck Millipore, Tullagreen, Ireland) and RHE were built as described before 21 .…”

Section: Methodsmentioning

confidence: 99%

Claudin-1 decrease impacts epidermal barrier function in atopic dermatitis lesions dose-dependently

Bergmann

Buenau

Vidal-y-Sy

et al. 2020

Sci Rep

Self Cite

106

View full text Add to dashboard Cite

The transmembrane protein claudin-1 is a major component of epidermal tight junctions (TJs), which create a dynamic paracellular barrier in the epidermis. Claudin-1 downregulation has been linked to atopic dermatitis (AD) pathogenesis but variable levels of claudin-1 have also been observed in healthy skin. To elucidate the impact of different levels of claudin-1 in healthy and diseased skin we determined claudin-1 levels in AD patients and controls and correlated them to TJ and skin barrier function. We observed a strikingly broad range of claudin-1 levels with stable TJ and overall skin barrier function in healthy and non-lesional skin. However, a significant decrease in TJ barrier function was detected in lesional AD skin where claudin-1 levels were further reduced. Investigations on reconstructed human epidermis expressing different levels of claudin-1 revealed that claudin-1 levels correlated with insideout and outside-in barrier function, with a higher coherence for smaller molecular tracers. Claudin-1 decrease induced keratinocyte-autonomous IL-1β expression and fostered inflammatory epidermal responses to non-pathogenic Staphylococci. In conclusion, claudin-1 decrease beyond a threshold level results in TJ and epidermal barrier function impairment and induces inflammation in human epidermis. Increasing claudin-1 levels might improve barrier function and decrease inflammation and therefore be a target for AD treatment. Tight junctions (TJs) are an important component of the complex epidermal barrier system. They are localized in the stratum granulosum (SG) of the epidermis and provide mechanical barrier function to ions and solutes of different molecular sizes 1-4. The transmembrane protein claudin-1 (Cldn-1) is a major component of TJs 5. It is also found outside of TJs in basal and suprabasal layers of the epidermis 2,5. Mice with a complete Cldn-1 knockout (KO) die at the first day of birth due to increased transepidermal water loss (TEWL) 5. They develop TJs leaky to a molecular tracer (Biotin-556) 5 , and a highly water permeable stratum corneum (SC) 6. Human subjects lacking Cldn-1 suffer from the Neonatal Ichthyosis-Sclerosing Cholangitis (NISCH) syndrome which includes an ichthyosiform skin phenotype 7. An archetypical disease of epidermal barrier dysfunction is atopic dermatitis (AD) 8. Cldn-1 single nucleotide polymorphisms were linked to AD in some cohorts 9-11 , but not in others 11,12. Using immunostaining-intensity measurements and western blot analyses, reduced Cldn-1 levels were found in lesional AD skin 13-16. For non-lesional skin, divergent observations were described. Some authors found decreased mRNA and immunointensity levels 10 , while others observed no alteration of Cldn-1 immunointensity and western-blot levels 14,16 .

show abstract

Section: Methodsmentioning

confidence: 99%

Claudin-1 decrease impacts epidermal barrier function in atopic dermatitis lesions dose-dependently

Bergmann

Buenau

Vidal-y-Sy

et al. 2020

Sci Rep

Self Cite

106

View full text Add to dashboard Cite

show abstract

“…Broadbent et al found the type of media used for differentiation and maintenance of airway epithelial cells impacts experimental results, including morphology, epithelial integrity, and response to viral infection ( Broadbent et al, 2020 ). Zorn-Kruppa et al also commented on KC characteristics altered by the propagation media ( Zorn-Kruppa et al, 2016 ). Key differences to note in these studies include: 1) primary KC were used, 2) comparison was with different propagation media than we studied (e.g., KGM2, DermaLife, and EpiLife; some of these media and supplements were backordered so we were unable to include them in our study), and 3) they used a protocol for KC differentiation (high calcium added to the culture media rather than differentiation media) which is not commonly used.…”

Section: Introductionmentioning

confidence: 99%

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

Moran¹,

Pope²,

Brewer³

et al. 2022

JID Innovations

View full text Add to dashboard Cite

“…Johanna Brandner gave a comprehensive introduction into tight junction biology, focusing on the role of claudin‐1 in atopic dermatitis . She also gave detailed methodological insights into an ex vivo wound healing model using porcine skin as well as cell culture methods for keratinocytes . The second keynote lecture of this session was given by Steffen Emmert, who provided detailed insight into DNA repair mechanisms in the skin and the contribution of Snap29 to epidermal differentiation and hair follicle formation .…”

mentioning

confidence: 99%

Matrix meets inflammation and DNA repair: Meeting report of the third ADF Round Table

Brandner

Emmert

Wolk

et al. 2017

Experimental Dermatology

View full text Add to dashboard Cite

Major cell biological parameters of keratinocytes are predetermined by culture medium and donor source

Cited by 7 publications

References 9 publications

Claudin-1 decrease impacts epidermal barrier function in atopic dermatitis lesions dose-dependently

Claudin-1 decrease impacts epidermal barrier function in atopic dermatitis lesions dose-dependently

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

Matrix meets inflammation and DNA repair: Meeting report of the third ADF Round Table

Contact Info

Product

Resources

About