Ker ct

Manufactured by American Type Culture Collection

Sourced in United States

The Ker-CT is a laboratory equipment product designed for cell culture applications. It functions as a specialized cell culture incubator, providing a controlled environment for the growth and maintenance of cells.

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Lab products found in correlation

9 protocols using ker ct

Keratinocyte Culture and Differentiation

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Neonatal human epidermal keratinocytes (HEKn, Life Technologies, Carlsbad, CA, USA) and immortalized human epidermal keratinocytes Ker-CT (ATCC, Manassas, VA, USA) were cultured in EpiLife medium with human keratinocyte growth supplements (HKGS, Life Technologies). HEK293T and H1299 cells were grown in DMEM medium (Lonza, Basel, Switzerland) with the addition of 10% FBS, 100 U penicillin, and 100 μg/mL streptomycin (Gibco, Life Technologies). Then, 2.5 × 10⁵ cells were transfected with 80 pmol of specific siRNAs (Supplementary Table 1) by Lipofectamine RNAiMAX Transfection Reagent (Invitrogen, Carlsbad, CA, USA) and collected 48 h post transfection. HEKn or Ker-CT cells were differentiated in vitro by adding 1.2 mM CaCl₂ to culture medium. Cells were collected at the indicated times or at 3 days of differentiation if not indicated otherwise. Further details for the generation of 3D organotypic skin model equivalents are described in Supplementary Material and Methods.

Smirnov A., Lena A.M., Cappello A., Panatta E., Anemona L., Bischetti S., Annicchiarico-Petruzzelli M., Mauriello A., Melino G, & Candi E. (2018). ZNF185 is a p63 target gene critical for epidermal differentiation and squamous cell carcinoma development. Oncogene, 38(10), 1625-1638.

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IL-6 Regulation of Epidermal Keratinocytes

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IL-6 knockout (KO) epidermis was collected from neonatal mice and isolated as previously described (13 (link)). Neonatal human epidermal keratinocytes (Ker-CT, ATCC CRL-4048 and nHEKs, ThermoFisher, Waltham, MA) were cultured according to manufactures protocol. Cells were treated with IL-6 (overnight for protein expression studies or 1 hour for signaling assays) or in combination with TGF-β1 (30 minutes) or specified otherwise. IL-6 neutralizing antibody (Biolegend, San Diego, CA) was used at 500ng/ml and incubated overnight followed by treatment. LLL-12 was used at .5μM for 1 hour.

Luckett-Chastain L.R., Cottrell M.L., Kawar B.M., Ihnat M.A, & Gallucci R.M. (2017). Interleukin (IL)-6 modulates transforming growth factor -β receptor I and II (TGF-βRI and II) function in epidermal keratinocytes. Experimental dermatology, 26(8), 697-704.

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Comparative Cell Line Analysis

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9e10, K562, Ker-CT, and A549 cells (ATCC)
were used in the study.

Pranauskaite E., Milkus V., Ritmejeris J., Zilionis R, & Mazutis L. (2024). Increasing Fluid Viscosity Ensures Consistent Single-Cell Encapsulation. Analytical Chemistry, 96(18), 6898-6905.

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Culturing Human Keratinocyte Cell Lines

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The HEK001 (CRL-2404, ATCC) human keratinocyte cell line, which was obtained from an older human subject, was cultured in keratinocyte-serum free medium (Gibco, Carlsbad, CA, USA, 17005-042) supplemented with human recombinant EGF (5 ng/mL), gentamicin (10 mg/mL), and 2 mM L-glutamine. The Ker-CT (CRL-4048, ATCC) human neonatal keratinocyte cell line was cultured in KGM Gold BulletKit medium (Lonza, Walkersville, MD, USA, 00192060). These cell lines were maintained in a humidified incubator at 37 °C with 5% CO₂. The protocols for each specific treatment are described in the figure legends.

Shen Z.Q., Chang C.Y., Yeh C.H., Lu C.K., Hung H.C., Wang T.W., Wu K.S., Tung C.Y, & Tsai T.F. (2024). Hesperetin activates CISD2 to attenuate senescence in human keratinocytes from an older person and rejuvenates naturally aged skin in mice. Journal of Biomedical Science, 31, 15.

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Airway Epithelial Cell Culture Protocol

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Insert transwells (Merck, Rahway, NJ, USA, MCHT12H48) were seeded with 200,000 human foreskin keratinocytes (Ker-CT, ATCC CRL-4048) on the transwells in 300 μL CnT-PR medium (CellnTec) in a 12-well format. After 48 h, the cultures were switched to CnT-PR-3D medium (CELLnTEC, Bern, Switzerland) for 24 h and then cultured in the air–liquid interface for 17 days. From day 12 to 17 of the air–liquid interphase culture, Th2 cytokines IL13 (10 ng/mL) and IL4 (10 ng/mL) (PeproTech, London, UK) were added [32 (link)]. Pharmacological treatments were applied from day 14 to 17 and consisted of 100 μM Olaparib, 100 μM FK-866 or their combination. The culture medium was refreshed every 2 days. On day 17, tissues were harvested for protein and gene expression analysis.

Arroyo A.B., Bernal-Carrión M., Cantón-Sandoval J., Cabas I., Corbalán-Vélez R., Martínez-Menchón T., Ferri B., Cayuela M.L., García-Moreno D, & Mulero V. (2023). NAMPT and PARylation Are Involved in the Pathogenesis of Atopic Dermatitis. International Journal of Molecular Sciences, 24(9), 7992.

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Human Keratinocyte Air-Liquid Culture

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Insert transwells (Merck, MCHT12H48) were seeded with 10⁵ human foreskin keratinocytes (Ker-CT, ATCC CRL-4048) on the transwells in 300 μL CnT-PR medium (CellnTec) in a 12 well format. After 48 hours, cultures were switched to CnT-PR-3D medium (CELLnTEC, Bern 3014, Switzerland) for 24 hours and then cultured at the air–liquid interface for 17 days. From day 12 to 17 of the air–liquid interphase culture, the Th17 cytokines IL17A (30 ng/mL) and IL22 (30 ng/mL) were added [60 (link)]. Pharmacological treatments were applied from day 14 to 17 and consisted of 100 μM apocynin, 100 μM FK-866, 100 μM olaparib, 10 μM NP and 1 μM ATRA. Culture medium was refreshed every 2 days. At day 17, the tissues were harvested for gene expression analysis.

Martínez-Morcillo F.J., Cantón-Sandoval J., Martínez-Navarro F.J., Cabas I., Martínez-Vicente I., Armistead J., Hatzold J., López-Muñoz A., Martínez-Menchón T., Corbalán-Vélez R., Lacal J., Hammerschmidt M., García-Borrón J.C., García-Ayala A., Cayuela M.L., Pérez-Oliva A.B., García-Moreno D, & Mulero V. (2021). NAMPT-derived NAD+ fuels PARP1 to promote skin inflammation through parthanatos cell death. PLoS Biology, 19(11), e3001455.

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Immortalized Keratinocyte Cultivation and EGF Supplementation

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Immortalized human keratinocytes derived from infant foreskin (KerCT®) were obtained from ATCC (VA). KerCT cells were cultured in the Keratinocyte Growth Medium (KGM, Lonza Bioscience, Switzerland) at 37°C, 5% CO₂. EGF (epidermal growth factor, AF‐100‐15, PeproTech, NJ) was used as a reagent during the incubation period (at a final concentration of 10 ng/mL).

Imafuku K., Iwata H., Natsuga K., Okumura M., Kobayashi Y., Kitahata H., Kubo A., Nagayama M, & Ujiie H. (2023). Zonula occludens‐1 distribution and barrier functions are affected by epithelial proliferation and turnover rates. Cell Proliferation, 56(9), e13441.

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Keratinocyte Differentiation and Th17 Modulation

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Insert transwells (Sigma-Aldrich MCHT12H48) were seeded with human foreskin keratinocytes (Ker-CT, ATCC CRL-4048) on the transwells in 300 μL CnT-PR medium (CellnTec) in a 12-well format. After 48 h, cultures were switched to CnTPR-3D medium (CELLnTEC) for 24 h and then cultured at the air-liquid interface for 17 days [30 (link)]. From day 12 to 17 of the air-liquid interphase culture, the Th17 cytokines IL17A (30 ng/mL) and IL22 (30 ng/mL) were added. Pharmacological treatments consisting of WZB117 at 100 μM, DPP4 at 100 μM, and all-trans retinoic acid (ATRA) at 1 μM (Table S2), were applied from day 14 to 17. The culture medium was refreshed every two days. On day 17, the tissues were harvested for gene expression analysis.

Fatás-Lalana B., Cantón-Sandoval J., Rodríguez-Ruiz L., Corbalán-Vélez R., Martínez-Menchón T., Pérez-Oliva A.B, & Mulero V. (2022). Impact of Comorbidities of Patients with Psoriasis on Phototherapy Responses. International Journal of Molecular Sciences, 23(17), 9508.

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Culturing Fibroblasts and Keratinocytes

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Normal human fibroblasts obtained from uninvolved skin of surgical specimens were cultured for 24 hours in Dulbecco's modified Eagle's medium (Life Technologies, Tokyo, Japan) containing 1.8 mM of calcium. An antibiotic-antimycotic solution (Sigma Aldrich, St. Luis, MO) and 10% fetal calf serum were supplied to the medium. Immortalized human keratinocytes (Ker-CT) purchased from ATCC (Manassas, VA) were cultured in keratinocyte growth medium (KGM-Gold, Lonza, Basel, Switzerland).
The recombinant human TNFα, IL-17A and IL-23 were purchased from PeproTech (Cranbury, NJ, 10602HNAE5), from Thermo Fisher Scientific (Waltham, MA, PHC9174) and from abcam (Cambridge, UK, ab106889), respectively.

Iwata H., Haga N, & Ujiie H. (2021). Possible role of epiregulin from dermal fibroblasts in the keratinocyte hyperproliferation of psoriasis. The Journal of dermatology, 48(9).

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