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HaCaT Cells

HaCaT cells are a spontaneously immortalized human keratinocyte cell line that retains the capacity for terminal differentiation.
These cells are widely used in skin biology and dermatological research as a model for human epidermal keratinocytes.
HaCaT cells exhibit a near-normal differentiation program and are a valuable tool for studying skin physiology, wound healing, and the effects of various agents on keratinocyte biology.
The availability of this well-characterized cell line has greatly facilitated the advancement of our understading of human skin structure and function.

Most cited protocols related to «HaCaT Cells»

1
Scratch Wound Healing Assay with hAdMSCs Conditioned Medium
We seeded the HaCaT cells in a 24-well plate at a concentration of 1x105 cells /well and we left them until they reached 80% of confluence [10 (link)]. We scratched cell cultures with a 200 μL sterile pipette tip [7 (link), 22 (link)] using the designed mold (see above for details) (S1 File). We then washed away detached cells with PBS (1X). Then, we added 1 mL of hAdMSCs conditioned medium, which was obtained from cells in the 4-6th passage according to the protocol established by Qazi et al. [7 (link)]. We made horizontal reference lines on the bottom of the plate with an ultrafine tip marker to have a grid for alignment to obtain the same field for each image acquisition run. Once we made the reference lines (approximately 3000 μm of distance), the plate was placed under a phase-contrast microscope using as guide the reference marks. We analyzed selected regions of interest using a Zeiss inverted microscope on a 10X objective with NA/0.25 every 2 hours for 28 hours. We determined the scratch area, wound coverage of total area, and average and standard deviation of the scratch width with the aid of our plugin. We calculated the rate of cell migration (RM) and the percentage of wound closure according to (Eq 1) and (Eq 2), respectively [28 (link)]:
RM=Wi−Wft;
WoundClosure%=(At=0−At=ΔtAt=0)x100%
Where Wi is the average of the initial wound width, Wf is the average of the final wound width both in μm and t is the time span of the assay in hours. Additionally, At = 0 is the initial wound area, At = Δt is the wound area after n hours of the initial scratch, both in μm2.
Suarez-Arnedo A., Torres Figueroa F., Clavijo C., Arbeláez P., Cruz J.C, & Muñoz-Camargo C. (2020). An image J plugin for the high throughput image analysis of in vitro scratch wound healing assays. PLoS ONE, 15(7), e0232565.
Publication 2020
Biological Assay Cell Culture Techniques Cells Culture Media, Conditioned Fungus, Filamentous HaCaT Cells Microscopy Microscopy, Phase-Contrast Migration, Cell Sterility, Reproductive Wounds
2
Alu Element Identification and Analysis
The Perl program “Alu_Mask” was written and used together with REPEATMASKER (http://www.repeatmasker.org/cgi-bin/WEBRepeatMasker) to define Alu elements. The Perl program “RNA_RNA_anneal” was generated to predict intermolecular duplexes between Alu elements within lncRNAs and proven or putative SMD targets. Duplexes were then validated using RNA structure 4.6 (http://rna.urmc.rochester.edu/rnastructure.html), which provides folding free energy changes. Human HeLa or HaCaT cells were transiently transfected with the specified plasmids or siRNAs as described1 (link). For mRNA half-life measurements, HeLa Tet-Off cells (Clontech) were utilized. If formaldehyde-crosslinked, cells were sonicated six times for 30 sec to facilitate lysis, and crosslinks were subsequently reversed by heating at 65°C for 45 min after IP. IP was performed as described1 (link). Protein was purified and Western blotting was performed as noted1 (link). RNA was purified from total, nuclear or cytoplasmic cell fractions or immunoprecipitated from total-cell lysates as reported1 (link). Poly(A)+ RNA was extracted from total-cell RNA using the Oligotex mRNA Mini Kit (Qiagen). RT-sqPCR and RT-qPCR were as described1 (link), except when RT was primed using oligo(dT)18 rather than random hexamers. The RNase protection assay employed the RPA III RNase Protection Assay Kit (Ambion) and uniformly labeled RNA probes that were generated by transcribing linearized pcDNA3.1(+)/Zeo_Chr11_66193000-66191383 in vitro using α-[P32]-UTP (Perkin Elmer) and the MAXIscript Kit (Ambion). Cells were visualized using a Nikon Eclipse TE2000-U inverted fluorescence microscope and, for phase microscopy, a 480-nm excitation spectra. Images were captured utilizing TILLVISION software (TILL Photonics). Scrape injury repair assays were essentially as published21,22. All data derive from at least three independently performed experiments that did not vary by more than the amount shown, and p values for all RT-sqPCR results were <0.05.
Gong C, & Maquat L.E. (2011). lncRNAs transactivate Staufen1-mediated mRNA decay by duplexing with 3'UTRs via Alu elements. Nature, 470(7333), 284-288.
Publication 2010
Alu Elements Biological Assay Cells Cytoplasm Endoribonucleases Formaldehyde HaCaT Cells HeLa Cells Homo sapiens Microscopy Microscopy, Fluorescence Oligonucleotides Plasmids Proteins RNA, Long Untranslated RNA, Messenger RNA, Polyadenylated RNA, Small Interfering Wound Healing
3
Immortalized Human Keratinocyte Culture Protocols
HaCaT cells, spontaneously immortalized human keratinocyte line [15 (link)], were kindly provided by Cell Line Service GmbH (Eppelheim, Germany) and cultured in 5% CO2 at 37°C in regular Dulbecco's Modified Eagle's Medium (DMEM) (Euroclone S.P.A., Milan, Italy) containing 1.8 mM Ca2+, or with DMEM (Gibco, Life Technologies, Carlsbad, CA, USA) at low concentration of Ca2+ (0.07 mM). Both media were supplemented with 10% heat-inactivated fetal bovine serum, glutamine (2 mM), penicillin (100 U⁄ml) (Euroclone), and streptomycin (100 mg⁄ml) (Euroclone). For all experiments, cells were seeded at a density of 5.7 × 103 cells⁄cm2 and cultured with DMEM at high or low Ca2+concentration for 6 or 14 days. The samples were labeled as follows: A6, cells cultured for 6 days with low Ca2+ concentration (0.07 mM) and tested when 80% confluent; A14, cells cultured for 14 days with low Ca2+ concentration (0.07 mM) and tested when overconfluent; C6, cells cultured for 6 days with high Ca2+ concentration (1.8 mM) and tested when 80% confluent; and C14, cells cultured for 14 days with high Ca2+ concentration (1.8 mM) and tested when overconfluent. The medium was changed every 2 days. A flow chart with details of the experimental protocol is reported in Figure 1.
Colombo I., Sangiovanni E., Maggio R., Mattozzi C., Zava S., Corbett Y., Fumagalli M., Carlino C., Corsetto P.A., Scaccabarozzi D., Calvieri S., Gismondi A., Taramelli D, & Dell'Agli M. (2017). HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes. Mediators of Inflammation, 2017, 7435621.
Publication 2017
Cell Lines Cells Fetal Bovine Serum Glutamine HaCaT Cells Homo sapiens Keratinocyte Penicillins Streptomycin
4
Multimodal Characterization of Cell Lines

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Publication 2009
Antibodies Calorimetry Cells Genes HaCaT Cells Immunoprecipitation Mouse Embryonic Stem Cells Mus Oligonucleotide Primers Real-Time Polymerase Chain Reaction RNA Interference Titrimetry Transfection
5
Culturing Human Keratinocytes for Vitamin D Research
Immortalized human keratinocytes (HaCaT) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with glucose, L-glutamine, pyridoxine hydrochloride (Cell Grow), 5% fetal bovine serum (Sigma-Aldrich, USA) and 1% penicillin/streptomycin/amphotericin antibiotic solution (Sigma) (Zbytek et al., 2003 (link)). Normal human keratinocytes (HEKn) were isolated from neonatal foreskin of African American donors using 2.5 UI/ml dispase and grown in keratinocyte basal medium (KBM) supplemented with keratinocytes growth factors (KGF) (Lonza) on collagen coated plates (Janjetovic et al., 2009 (link)). For experiments cells in their third passage were used. Prior to treatment with vitamin D3 metabolites, HaCaT cells were serum deprived for 24 h and further incubated in DMEM medium containing 5% charcoal treated serum (HyClone). For HEKn cells, 0.5% BSA solution was used when cells were treated with vitamin D3 derivatives.
JANJETOVIC Z., TUCKEY R.C., NGUYEN M.N., THORPE EM J.R, & SLOMINSKI A.T. (2010). 20,23-Dihydroxyvitamin D3, Novel P450scc Product, Stimulates Differentiation and Inhibits Proliferation and NF-κB Activity in Human Keratinocytes. Journal of cellular physiology, 223(1), 36.
Publication 2009
African American Amphotericin Cells Charcoal Cholecalciferol Collagen derivatives dispase Donors Eagle Fetal Bovine Serum FGF7 protein, human Foreskin Glucose Glutamine HaCaT Cells Homo sapiens Infant, Newborn Keratinocyte Penicillins Pyridoxine Hydrochloride Serum Streptomycin

Most recents protocols related to «HaCaT Cells»

1
Peptide Reduces TCDD-Induced ROS in HaCaT Cells
Not available on PMC !

Example 4

HaCaT cells, human keratinocyte cells, were seeded on a 6-well plate at a density of 3×105 cells/well and cultured overnight. Subsequently, 10 nM of TCDD and 50 μM of the peptide consisting of an amino acid sequence of SEQ ID NO: 1, 2, or 3 were added to the culture medium. After 30 minutes of reaction, the cells were treated for 24 hours and further treated with DCFH-DA for 30 minutes. Then, the cells were collected and subjected to FACS analysis to observe changes of average FL1 values, and the results are shown in FIGS. 4A to 4C.

As shown in FIGS. 4A to 4C, the peptide consisting of an amino acid sequence of SEQ ID NO: 1, 2, or 3 reduced ROS levels increased by TCDD in cells.

US11859016B2. Peptide having cytoprotective effect against environmental pollutant and use thereof (2024-01-02). CAREGEN CO., LTD. [KR]. Inventors: Yong Ji Chung [KR], Eun Mi Kim [KR], Eung-ji Lee [KR].
Patent 2024
Amino Acid Sequence Cells Culture Media diacetyldichlorofluorescein Figs HaCaT Cells Homo sapiens Keratinocyte Peptides Tetrachlorodibenzodioxin
2
Inhibition of TCDD Cell Uptake by Peptides
Not available on PMC !

Example 3

HaCaT cells, human keratinocyte cells, were seeded on a 6-well plate at a density of 3×105 cells/well and cultured overnight. Subsequently, 50 nM of TCDD and 50 μM of the peptide consisting of an amino acid sequence of SEQ ID NO: 1, 2, or 3 were added to the culture medium. After 30 minutes of reaction, the cells were treated for 5 minutes and immobilized with 4% paraformaldehyde for 30 minutes. Then, after washing three times, the cells were reacted with 0.5% Triton X-100 for 15 minutes and washed three times. Subsequently, the cells were blocked with 3% BSA for 1 hour and reacted with a primary antibody against TCDD conjugated with fluorescein isothiocyanate (FITC) (1:100) at 4° C. overnight. The cells were stained and mounted with 4,6-diamidino-2-phenylindole (DAPI) and observed with a fluorescence microscope. The results are shown in FIGS. 3A to 3C.

As shown in FIGS. 3A to 3C, the peptide consisting of an amino acid sequence of SEQ ID NO: 1, 2, or 3 inhibited introduction of TCDD into cells.

US11859016B2. Peptide having cytoprotective effect against environmental pollutant and use thereof (2024-01-02). CAREGEN CO., LTD. [KR]. Inventors: Yong Ji Chung [KR], Eun Mi Kim [KR], Eung-ji Lee [KR].
Patent 2024
Amino Acid Sequence Cells Culture Media Figs Fluorescein HaCaT Cells Homo sapiens Immunoglobulins isothiocyanate Keratinocyte Microscopy, Fluorescence paraform Peptides Tetrachlorodibenzodioxin Triton X-100
3
HaCaT Cell Culture and Cytotoxicity Assay
Human
Keratinocyte skin cells (CVCL_0038 and CLS: 300493) were acquired
from Cell Lines Services (CLS, Germany). For HaCaT culture, RPMI-1640
media (Serana, Germany) was used complemented with 1% MEM NEAA, 1%
L-glutamine, 1% penicillin streptomycin antibiotic, and 10% FBS in
75 cm2 culture flasks in a humidified 37 °C incubator
with 5% CO2. To detach the adherent HaCaT cells to be used
in cytotoxicity assays, trypsin was added to cells. HaCaT cells were
then seeded in 96-well plates and incubated at 37 °C for 24 h
or until a monolayer was observed.45 (link)
Abdelnasir S., Mungroo M.R., Chew J., Siddiqui R., Khan N.A., Ahmad I., Shahabuddin S, & Anwar A. (2023). Applications of Polyaniline-Based Molybdenum Disulfide Nanoparticles against Brain-Eating Amoebae. ACS Omega, 8(9), 8237-8247.
Publication 2023
Biological Assay Cell Lines Cells Complement C1 Cytotoxin Glutamine HaCaT Cells Penicillins Skin Streptomycin Trypsin
4
NB-UVB-Induced Cytotoxicity Mitigation

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Publication 2023
Biological Assay Cells Cell Survival HaCaT Cells Radiotherapy
5
NB-UVB Irradiation and DSE Pretreatment

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Publication 2023
Biological Assay Cells Cell Survival HaCaT Cells

Top products related to «HaCaT Cells»

1
Fetal bovine serum (fbs) by Thermo Fisher Scientific
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
2
Dulbecco modified eagle medium (dmem) by Thermo Fisher Scientific
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DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture medium formulated to support the growth and maintenance of a variety of cell types, including mammalian cells. It provides essential nutrients, amino acids, vitamins, and other components necessary for cell proliferation and survival in an in vitro environment.
3
Penicillin by Thermo Fisher Scientific
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Penicillin is a type of antibiotic used in laboratory settings. It is a broad-spectrum antimicrobial agent effective against a variety of bacteria. Penicillin functions by disrupting the bacterial cell wall, leading to cell death.
4
Streptomycin by Thermo Fisher Scientific
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Streptomycin is a broad-spectrum antibiotic used in laboratory settings. It functions as a protein synthesis inhibitor, targeting the 30S subunit of bacterial ribosomes, which plays a crucial role in the translation of genetic information into proteins. Streptomycin is commonly used in microbiological research and applications that require selective inhibition of bacterial growth.
5
Penicillin streptomycin by Thermo Fisher Scientific
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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
6
Hacat cells by American Type Culture Collection
Sourced in United States
HaCaT cells are a spontaneously immortalized human keratinocyte cell line. They are derived from normal human skin and exhibit characteristics of basal keratinocytes. HaCaT cells are a widely used in vitro model for the study of keratinocyte biology and skin-related research.
7
Lipofectamine 2000 by Thermo Fisher Scientific
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Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.
8
Trizol reagent by Thermo Fisher Scientific
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TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
9
Methyl thiazolyl tetrazolium (mtt) by Merck Group
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MTT is a colorimetric assay used to measure cell metabolic activity. It is a lab equipment product developed by Merck Group. MTT is a tetrazolium dye that is reduced by metabolically active cells, producing a colored formazan product that can be quantified spectrophotometrically.
10
Dulbecco modified eagle medium (dmem) by Merck Group
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DMEM (Dulbecco's Modified Eagle's Medium) is a commonly used cell culture medium formulated to support the growth and maintenance of various cell lines. It provides a balanced salt solution and essential nutrients required for cell proliferation. DMEM is suitable for use in a wide range of cell culture applications.

More about "HaCaT Cells"

HaCaT cells are a widely-used human keratinocyte cell line that exhibits a near-normal differentiation program, making them a valuable model for studying skin physiology, wound healing, and the effects of various agents on keratinocyte biology.
These spontaneously immortalized cells are derived from human skin and retain the capacity for terminal differentiation, making them a reliable and well-characterized tool for dermatological research.
HaCaT cells are commonly cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with fetal bovine serum (FBS) and antibiotics like penicillin and streptomycin.
This serum-containing medium provides the necessary nutrients and growth factors to support the proliferation and differentiation of the HaCaT cells.
Researchers often utilize transfection reagents like Lipofectamine 2000 to introduce genetic material, such as plasmids or small interfering RNAs (siRNAs), into HaCaT cells.
This allows for the study of gene expression and the effects of specific gene manipulation on keratinocyte biology.
Additionally, techniques like the MTT assay are commonly employed to assess cell viability and proliferation in HaCaT cell cultures.
The availability of the well-characterized HaCaT cell line has greatly facilitated the advancement of our understanding of human skin structure and function.
Researchers can leverage the power of PubCompare.ai, an AI-driven platform, to effortlessly locate protocols from literature, preprints, and patents, and utilize its intelligent comparisons to identify the optimal protocols and products for their HaCaT cell studies.
This cutting-edge technology simplifies the research process and enables more effective and streamlined experimentation.