Nheks

Manufactured by ScienCell

Sourced in United States

NHEKs are primary human epidermal keratinocytes derived from neonatal foreskin tissue. They are a key component of the epidermal layer of human skin and play a crucial role in maintaining skin barrier function.

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Lab products found in correlation

3 protocols using nheks

HPV16 E7 Gene Cloning and Expression

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HPV16-containing plasmid DNA (ATCC:45113D) was used as the template for PCR amplification and cloning of the HPV16 E7 gene into vector pEGFP-C1 following protocols described previously. NHEKs (ScienCell, Carlsbad, CA, United States) were cultured in 6-well plates using EpiLife culture medium (Gibco, United States) with 10% fetal bovine serum (Gibco, United States) and Human Keratinocyte Growth Supplement (Gibco, United States). NHEKs at 80% confluence were transfected with pEGFP-16E7 and pEGFP-C1 using Lipofectamine 3000 reagent (Invitrogen, Carlsbad, CA, United States). Six hours after Transfection, culture medium was replaced with fresh culture medium.

Hua C., Zhu J., Zhang B., Sun S., Song Y., van der Veen S, & Cheng H. (2020). Digital RNA Sequencing of Human Epidermal Keratinocytes Carrying Human Papillomavirus Type 16 E7. Frontiers in Genetics, 11, 819.

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Mimicking Abnormal Keratinocyte Proliferation

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Normal human epidermal keratinocytes (NHEKs) were obtained from ScienCell Research Laboratories, Inc. NHEKs were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Thermo Fisher Scientific, Inc.), 100 U/ml penicillin (Thermo Fisher Scientific, Inc.) and 100 mg/ml streptomycin (Thermo Fisher Scientific, Inc.), and maintained in an incubator at 37°C in the presence of 5% CO₂. NHEKs were treated with 10 ng/ml M5 [IL-1α, IL-17A, IL-22, oncostatin M (OSM) and TNF-α; PeproTech China] for 24 h at 37°C to mimic abnormal proliferation and differentiation of keratinocytes (9 (link)).

Ding Z., Liu J., Qian H., Wu L, & Lv M. (2021). Cinnamaldehyde inhibits psoriasis-like inflammation by suppressing proliferation and inflammatory response of keratinocytes via inhibition of NF-κB and JNK signaling pathways. Molecular Medicine Reports, 24(3), 638.

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Sourcing and Culturing Cell Lines

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Neonatal human dermal fibroblasts (NDFs) were purchased from ATCC (ATCC, PCS-201-010, Manassas, VA, USA). Human iPSC endothelial cells were purchased from FUJIFILM (FUJIFILM Cellular Dynamics, R1022, WI, USA) and maintained in iCell endothelial cells medium (Supplemental Table 1). Neonatal human epidermal keratinocytes (NHEKs) were purchased from Sciencell (Sciencell,2100, Carlsbad, CA, USA). NDFs were maintained in MEM medium supplemented with 5 % FBS (Thermo Fisher Scientific, Waltham, MA, USA). NHEKs were maintained in keratinocyte growth medium (KGM) purchased from Sciencell (Sciencell, 2101, Carlsbad, CA, USA). Human placental microvascular pericytes were purchased from agio-protemomie(Angio-proteomie, cAP-0029, Boston, MA, USA) and cultured in MEM medium supplemented with 5 % FBS (Thermo Fisher Scientific, Waltham, MA, USA). All the cells were maintained at 37 °C under a 5 % CO₂ atmosphere. Cells of passage number 3-4 were used. All the cultures were routinely monitored for mycoplasma contamination using MycoAlert PLUS mycoplasma detection kit (Lonza, Walkersville, MD, USA).

Liu X., Michael S., Bharti K., Ferrer M, & Song M.J. (2020). A Biofabricated Vascularized Skin Model of Atopic Dermatitis for Preclinical Studies. Biofabrication, 12(3), 035002.

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