Hacat cells

Manufactured by Merck Group

Sourced in United States, Germany

HaCaT cells are a spontaneously transformed, immortalized human keratinocyte cell line. They are commonly used as an in vitro model for studying skin biology, wound healing, and other dermatological processes.

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Lab products found in correlation

Streptomycin, by Merck Group (3 mentions) Penicillin, by Merck Group (3 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Heracell 240i, by Thermo Fisher Scientific (1 mentions) Thp 1, by American Type Culture Collection (1 mentions) L glutamine, by Merck Group (1 mentions) Rpmi 1640, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions) Glutamine, by Merck Group (1 mentions) Streptomycin, by Avantor (1 mentions) Penicillin g, by Avantor (1 mentions) Hacat cell, by Cytion (1 mentions) Hacat, by CLS Cell Lines Service (1 mentions) Mcf 7, by CLS Cell Lines Service (1 mentions) Mcf 7, by Merck Group (1 mentions) Rpmi 1640 medium, by Lonza (1 mentions)

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HaCaT (16 citations)

5 protocols using hacat cells

Growth of Human Cell Lines

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Human keratinocytes cell lines from adult skin (HaCaT cells) purchased by Cell Line Service (CLS Eppelheim, Baden-Württemberg, Germany) were grown in Dulbecco’s modified Eagle’s medium (DMEM, Merck KGaA, Darmstadt, Germany), supplemented with 4.5 g/L glucose, 2 mM L-glutamine and 10% heat-inactivated fetal bovine serum (FBS) (Merck KGaA, Darmstadt, Germany).
The human monocytic leukemia cell line (THP-1) was purchased from the ATCC (Manassas, VA, USA) and maintained in RPMI 1640 (Merck KGaA, Darmstadt, Germany) added with 0.05 mM of 2-mercaptoethanol, 10% of FBS, glutamine (2 mM), penicillin (100 U/mL), and streptomycin (100 mg/mL) (Merck KGaA, Darmstadt, Germany). Both cells were cultured at 37 °C in a 5% CO₂ humidified incubator (HeraCell 240i, Thermo-Fisher Scientific, Waltham, MA, USA).

Marinelli L., Cacciatore I., Costantini E., Dimmito M.P., Serra F., Di Stefano A, & Reale M. (2022). Wound-Healing Promotion and Anti-Inflammatory Properties of Carvacrol Prodrugs/Hyaluronic Acid Formulations. Pharmaceutics, 14(7), 1468.

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Scratch Wound Healing Assay with HaCaT Cells

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The human keratinocyte (HaCaT) cells (obtained from the American Type Culture Collection (ATCC), USA) were seeded in 12-well plates (1 × 10⁵ cells/well) and incubated for 24 h in a humidified 5% CO₂ incubator (37 °C) in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS), 100 units/mL of penicillin and 100 µg/mL of streptomycin (Merck Millipore, Burlington, MA, USA). Then, a scratch at the bottom of each well was made with tips, and different Cyclopia sp. extracts were added to the cells and incubated for up to 96 h. After treatment, the cells were washed and analysed under a microscope (magnification × 100) (Leica, Switzerland). All samples were compared to the untreated control cells.

Hering A., Stefanowicz-Hajduk J., Gucwa M., Wielgomas B, & Ochocka J.R. (2023). Photoprotection and Antiaging Activity of Extracts from Honeybush (Cyclopia sp.)—In Vitro Wound Healing and Inhibition of the Skin Extracellular Matrix Enzymes: Tyrosinase, Collagenase, Elastase and Hyaluronidase. Pharmaceutics, 15(5), 1542.

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Culturing and Treating Human Keratinocytes

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Human keratinocytes (HaCaT cells) were obtained from Cell Lines Service (CLS;
Eppelheim, Germany). Cells were maintained in Dulbecco’s modified Eagle’s medium
(DMEM; Gibco BRL, Carlsbad, CA, USA), which was supplemented with 10%
heated-inactivated fetal bovine serum (FBS; Gibco BRL), 100 U/mL penicillin G,
and 100 mg/L streptomycin (both from Amresco, Solon, OH, USA), and cells were
cultured in a humidified cell culture incubator containing 5% CO₂ and
95% air at 37°C. For the treatment of IL-6, HaCaT cells were incubated in DMEM
containing IL-6 (Sigma-Aldrich, St. Louis, MO, USA) for 2 h.

Dong H., Jiang W., Chen H., Jiang S., Zang Y, & Yu B. (2018). MicroRNA-145 attenuates IL-6-induced enhancements of sensitivity to UVB irradiation by suppressing MyD88 in HaCaT cells. International Journal of Immunopathology and Pharmacology, 32, 2058738418795940.

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Cell Culture and Cytotoxicity Evaluation

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MCF-7, 5637 and HaCaT cells were obtained from CLS Cell Lines Service (Eppelheim, Germany). Cells were cultured in RPMI 1640 medium (BioWhittaker, Lonza, Belgium) supplemented with 8% fetal bovine serum (Sigma Aldrich, Germany) and antibiotics (100 U/mL penicillin/100 µg/mL streptomycin; Sigma Aldrich, Germany) at 95% humidity, 5% CO₂ and 37.5 ◦C. MCF-7, 5367 and HaCaT cells were sub-cultured twice a week and regularly tested for mycoplasma contamination. Cell viability (cytotoxicity) of the test samples was investigated on the cell lines using the neutral red uptake (NRU) assay.

Elhawary S., EL-Hefnawy H., Mokhtar F.A., Sobeh M., Mostafa E., Osman S, & El-Raey M. (2020). Green Synthesis of Silver Nanoparticles Using Extract of Jasminum officinal L. Leaves and Evaluation of Cytotoxic Activity Towards Bladder (5637) and Breast Cancer (MCF-7) Cell Lines. International Journal of Nanomedicine, 15, 9771-9781.

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Epithelial Cell Lines for HPV Research

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In the present study we solely used epithelial cells. The HPV-negative epithelial cell lines used were: HEK 293T (transformed human embryonic kidney cells), HaCaT (non-transformed immortalized human skin keratinocytes), HEKn (human epidermal keratinocytes), C33A (human cervical cancer cells), and H1299 (human lung non-small cancer cells). The HPV16-positive cell lines used were: CaSki (human cervical cancer cells) and SiHa (human cervical cancer cells). All cell lines were purchased from ATCC (Manassas, VA, USA) except for HaCaT cells which were a kind gift form Prof. Enzo Di Iorio (University of Padua, Italy) and HEKn which were purchased from Sigma-Aldrich (St. Louis, MO, USA). All cell lines were cultured in Dulbecco’s Modified Eagle’s medium (DMEM, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Waltham, MA, USA) except for HEKn which were cultured in Keratinocyte Growth Medium (Sigma-Aldrich, St. Louis, MO, USA). Cells were grown at 37 °C with 5% CO₂ and were tested monthly for mycoplasma contaminations. We did not regularly use Pen/Strep in cell culture media.

Messa L., Celegato M., Bertagnin C., Mercorelli B., Alvisi G., Banks L., Palù G, & Loregian A. (2021). The Dimeric Form of HPV16 E6 Is Crucial to Drive YAP/TAZ Upregulation through the Targeting of hScrib. Cancers, 13(16), 4083.

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