Nheks

Manufactured by Merck Group

NHEKs are a type of primary human epidermal keratinocytes, which are the predominant cell type in the epidermis. They are used in cell culture and research applications to study skin biology, wound healing, and related processes.

Automatically generated - may contain errors

Lab products found in correlation

5 protocols using nheks

Epidermal Keratinocyte Differentiation and Bacterial Toxin Response

Check if the same lab product or an alternative is used in the 5 most similar protocols

Normal neonatal human primary epidermal keratinocytes (NHEKs) (Thermo Fisher Scientific, Waltham, Mass) were cultured in Epilife complete medium containing 60 mM CaCl₂ (Thermo Fisher Scientific) supplemented with 1× human keratinocyte growth supplement (Thermo Fisher Scientific) and a 1× antibiotic-antimycotic (100 U/mL of penicillin, 100 U/mL of streptomycin, and 250 ng/mL of amphotericin B [Thermo Fisher Scientific]) at 37°C and 5% CO₂. NHEKs were used only for experiments between passages 3 to 5. NHEKs were grown to about 90% to 100% confluency followed by differentiation in Epilife complete medium with 2 mM CaCl₂ for 72 hours. For bacterial supernatant treatments, differentiated NHEKs were treated with sterile-filtered bacterial supernatant at 5% (vol) in Epilife medium for 8 hours and then harvested for RNA extraction. For treatment with EcpA, the purified enzyme (final concentration 2.5 μg/mL) was added to the culture medium along with 5 mM of L-cysteine over the course of either 8 hours for RNA extraction or 20 hours for protein extraction. Some NHEKs were treated with cycloheximide (final concentration 20 μg/mL) (Sigma-Aldrich, St Louis, Mo) 2 hours before treatment with EcpA and for the entire duration of the treatment.

Cau L., Williams M.R., Butcher A.M., Nakatsuji T., Kavanaugh J.S., Cheng J.Y., Shafiq F., Higbee K., Hata T.R., Horswill A.R, & Gallo R.L. (2020). Staphylococcus epidermidis protease EcpA can be a deleterious component of the skin microbiome in atopic dermatitis. The Journal of allergy and clinical immunology, 147(3), 955-966.e16.

+ Open protocol

+ Expand

Cell Culture Protocols for HeLa, U2OS, and NHEK

Check if the same lab product or an alternative is used in the 5 most similar protocols

HeLa cells (CRUK-CI Biorepository and Cell Services Core) were cultured in DMEM (Sigma, D6429) supplemented with 10 % heat inactivated FBS. U2OS AID-DIvA cells were cultured in DMEM supplemented with 10 % heat inactivated FBS and 800 μg/mL G418 Sulfate (Gibco, 10131). To induce nuclear localization of AsiSI, AID-DIvA cells were treated with 300 nM 4OHT (Sigma, H7904) for 4 h. Normal human epidermal keratinocytes (NHEK), pooled from multiple donors, were purchased from Thermofisher (A13401) and cultured in EpiLife medium (Thermofisher, M-EPI-500-CA) supplemented with human keratinocyte growth supplement (HKGS) (Thermofisher, S-001-5). NHEKs were detached using accutase (Sigma, A6964). HeLa and U2OS AID-DIvA cells were STR genotyped and mycoplasma tested. NHEK cells were obtained from Thermofisher and were certified as mycoplasma negative.

Lensing S.V., Marsico G., Hänsel-Hertsch R., Lam E.Y., Tannahill D, & Balasubramanian S. (2016). DSBCapture: in situ capture and direct sequencing of dsDNA breaks. Nature methods, 13(10), 855-857.

+ Open protocol

+ Expand

Keratinocyte Response to Uremic Toxins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Normal human epidermal keratinocytes (NHEKs; Lonza, Basel, Switzerland) were incubated with keratinocyte growth medium (Lonza) and serially subcultured for 5-10 passages. IS and PC were purchased from Sigma-Aldrich (St. Louis, Missouri, USA), and serum samples were obtained from CKD patients and healthy controls. IS and PC were diluted in phosphate-buffered saline and methanol, respectively, to concentrations of 10, 50, 100, and 250 mg/ mL. NHEKs were stimulated with different concentrations of IS and PC and with 4% pooled serum samples. NHEKs cultured with 4% bovine serum albumin were used as controls. In further experiments, NHEKs were additionally pretreated with soybean trypsin inhibitor (Sigma-Aldrich) at a dilution ratio of 1:10 in keratinocyte growth medium for each experimental treatment (differing concentrations of IS and PC) to evaluate its inhibitory effect on PAR-2 expression. All experiments were performed in triplicate.

Kim S.J., Zhang X., Cho S.B., Kim C.H., Park H.C, & Moon S.J. (2021). Uremic solutes of indoxyl sulfate and p-cresol enhance protease-activated receptor-2 expression in vitro and in vivo in keratinocytes. Human & experimental toxicology, 40(1).

+ Open protocol

+ Expand

Cell Culture Protocols for HeLa, U2OS, and NHEK

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lensing S.V., Marsico G., Hänsel-Hertsch R., Lam E.Y., Tannahill D, & Balasubramanian S. (2016). DSBCapture: in situ capture and direct sequencing of dsDNA breaks. Nature methods, 13(10), 855-857.

+ Open protocol

+ Expand

Evaluating Capsaicin's Effects on NHEKs

Check if the same lab product or an alternative is used in the 5 most similar protocols

We purchased normal human epidermal keratinocytes (NHEKs) from EpiLife (Cascade Biologics, Portland, OR, USA). Cells were cultured in basal keratinocyte growth medium (EpiLife), supplemented with human keratinocyte growth supplement (HKGS) and antibiotics (100 U/ml penicillin and 100 mg/ml streptomycin) in a 5% CO₂ incubator. Passages 2~9 were used for all experiments. NHEKs (1×10⁴ cells/well) were seeded into 96 well plates. After application of capsaicin and capsazepine (Sigma-Aldrich, St. Louis, MO, USA) at various concentrations, cell viability was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (Sigma-Aldrich) assay according to the manufacturer’s instructions. NHEKs at 70% confluence were stimulated with capsaicin (Sigma-Aldrich). The expression of TRPV1, LL37, KLK-5, TNF-α, VEGF, IL-1α, IL-1β, IL-8, and PAR2 was evaluated at times ranging from 0 to 12 h and at various concentrations of capsaicin (0.1, 1, 5, 10, and 20 µM). Regulation of these mediators induced by capsaicin (20 µM) was also evaluated after the application of a specific TRPV1 antagonist, capsazepine (Sigma-Aldrich) at various concentrations (1, 5, 10, and 20 µM). capsazepine was added 30 min before capsaicin stimulation and incubated for the indicated times.

Kim H.B., Na E.Y., Yun S.J, & Lee J.B. (2022). The Effect of Capsaicin on Neuroinflammatory Mediators of Rosacea. Annals of Dermatology, 34(4), 261-269.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!