hacat cell line characteristics

It was shown that immortalized, non-tumor-derived cell lines appear to behave very similarly to primary cells, keeping major specific characteristics, at least in early passages (Sprenger et al. J Tissue Cult Meth. Cells of the MCF-10A, PZ-HPV-7, HepG2, and MDA-MB-231 cell lines were purchased from the American Type Culture Col- Nat Rev Genet. Bondi CA, Marks JL, Wroblewski LB, Raatikainen HS, Lenox SR, Gebhardt KE. Fuchs E, Raghavan S. Getting under the skin of epidermal morphogenesis. HaCaT cells are utilized for their high capacity to differentiate and proliferate in vitro. Found insideHuman skin cancers, the most common type of tumors, represent a significant health burden. The deadliest is unquestionably melanoma. Rezaei-Tavirani M, Mansouri V, Mahdavi SM, Valizadeh R, Rostami-Nejad M, Zali MR. (2014) found dysregulation of the isoform Long of 14-3-3 protein beta/alpha in HaCaT cells treated with arsenic trioxide (8 passages). Proteins matched by one unique peptide were also considered when could be identified in all replicates. Anderson L. Six decades searching for meaning in the proteome. Luo W, Brouwer C. Pathview: an R/Bioconductor package for pathway-based data integration and visualization. Udensi UK, Tackett AJ, Byrum S, Avaritt NL, Sengupta D, Moreland LW, Tchounwou PB, Isokpehi RD. To construct the tissue engineering seed cell (HaCaT cell line) with stable expression of the human epidermal growth factor (EGF), and analyze the changes of its biological characteristics. 2008;128(5):1212–9. Article  Here, we employed human HaCaT cells as a model system to identify cellular proteins that accompany SDS-induced changes due to subtoxic concentration based on a proteomic approach. Regulatory changes of protein contents or enzyme activities are one of the biochemical features of diseases. The HaCaT epidermal epithelial cell line was established by Norbert E. Fusenig of the German Cancer Research Center (39), and culture conditions were the same as those previously described (39). The role of barrier genes in epidermal malignancy. The mRNA expression levels of the obtained HaCaT-EGF cell were more than 100 times higher than the level of ordinary HaCaT cell. 2009). The HaCaT cells were maintained in RPMI-1640 (Hyclone) supplemented with 10% fetal bovine serum (FBS, Hyclone), 1% antibiotics penicillin-streptomycin (P/S, Hyclone) and were . Toxicological interactions among arsenic, cadmium, chromium, and lead in human keratinocytes. The cell biology collection includes more than 4,000 continuous cell lines available by species, tissue/disease types, and signaling pathways. Cell Metab. in proinflammatory cytokine- and vehicle-treated HaCaT keratinocytes was performed. CAS  Methods Mol Biol. By NR uptake, it was established that there is a strong correlation between in vitro cytotoxicity in a human keratinocyte line (HaCaT) and in vivo human data for a homologous series of anionic surfactants (Wilhelm et al. Hoh A, Maier K, Dreher RM. Comparative quantitation of proteome alterations induced by aging or immortalization in primary human fibroblasts and keratinocytes for clinical applications. Although cell death by apoptosis has been recognized as an important control mechanism in the maintenance of tissue homeostasis and in the elimination of cells with damaged DNA, information on the induction and characteristics of apoptosis in keratinocytes is rather scarce. Google Scholar. Methods: HaCaT-HPV16-E2 cells were generated by transduction of HaCaT cell line with a lentiviral vector. 1994;32:5–8. Exposure of SDS to mammals results in physical and biochemical effects: skin irritation, hyperplasia, alteration of serum lipid composition, damage of cells, and decrease in cell proliferation (Lindberg et al. Gene Ontology (GO) overrepresentation analysis of upregulated proteins during the SDS treatment visualized using REVIGO (http://revigo.irb.hr). Nat Methods. 2.3 Preparation of coumarin 6-loaded DOC-mediated LS (DOC-LS) and conventional LS Ada Tse. It was shown that HaCaT cell line is characterized by a dose-dependent decrease in viability when exposed to sodium dodecyl sulphate. Sciacovelli M, Guzzo G, Morello V, Frezza C, Zheng L, Nannini N, Calabrese F, Laudiero G, Esposito F, Landriscina M, Defilippi P, Bernardi P, Rasola A. Treatment of HaCaT cells with 2,4‐dinitrochlorobenzene (DNCB) and benzalkonium . Changes in viability of immortalized HaCaT keratinocytes when exposed to different doses of SDS for 48 h were studied using light microscopy, MTT assay, and NR uptake test. 1996;288(11):684–90. 2001). They were incubated in a humidified incubator at 37 °C in 5% CO2 for a period of 48 h. Cytotoxicity in HaCaT cells following exposure to SDS was determined by measuring the viability of cells using MTT (tetrazolium salts) assay (Mossmann 1983) or NR assay (Borenfreund and Puerner 1984). AVL interpreted the proteomic data. The examination of HaCaT keratinocytes proteome was performed by Progenesis LC-MS software (Nonlinear Dynamics Ltd.), where two peak lists (separately for unexposed and exposed to SDS cells) were generated and exported for a Mascot MS/MS search. Google Scholar. Part of In amounts of 2 to 5%, SDS can cause sensitizing reactions, increase the transepidermal water loss of the stratum corneum, and result in mild skin inflammation affecting keratinocytes (Törmä et al. LC-MS/MS analyses showed that HaCaT cells (Additional file 1: Table S1 and Additional file 2: Table S2) contain several members of the 14-3-3 family of proteins including 1433S, which regulates protein synthesis and epithelial cell growth. GO enrichment analysis (Fig. 4 Ci ) but did form tumors in four of five animals after 90 days (not shown). Shi et al. This hypothesis is further supported by the remarkable decrease in cell viability of HaCaT cells upon stimulation with TNF-α and IFN-γ and increased expression of IL-33 from damaged HaCaT cells. 1) and were in accordance with microscopic observations (Fig. CAS  Biological characteristics; Gene transfection; HaCaT; Tissue engineering. Storage = 2ºC to 8ºC. Quantitative assessment of primary skin irritants in vitro in a cytotoxicity model: comparison with in vivo human irritation tests. 2009;8:32. For example, Rezaei-Tavirani et al. The pair of HaCaT samples (175 μg of protein) for each cell type (untreated and SDS-exposed) was processed in-solution digestion as described previously (Petushkova et al. 2013;32(3):269–76. HaCaT cells showed dissimilar responses to cytokine stimulation compared with primary KC in the expression of cornified envelope transcripts, suggesting that HaCaT cells are not a suitable alternative to primary KC (Seo et al., 2012). Google Scholar. Petushkova, N.A., Rusanov, A.L., Pyatnitskiy, M.A. Resulting lists of enriched Gene Ontology categories were visualized with the help of REViGO (Supek et al. This approach can provide candidate protein biomarkers related to the diseases. 1987;1(4):537–46. Authoritative and easily accessible, Epithelial Cell Culture Protocols, Second Edition will serve outstanding investigators with the best possible information for the advancement of biomedical science. Dickson FM, Lawrence JN, Benford DJ. ALR did cell culture experiments, interpreted proteomic data, and was a one of two major contributors in writing the manuscript. Divided into three convenient parts, the volume begins with a number of basic in vitro migration assays including measurements of wound healing, cell scattering, invasion, and chemotaxis, as well as more complex measurements of ... Published by Elsevier B.V. All rights reserved. Several studies on the effects of UV radiation have been included at the beginning of this volume. Even the role of long wavelength UV (UVA) has become a focus of research activities. The colony of the HaCaT-EGF cells was more focused and tight compared to the empty vector transfected HaCaT cells and normal HaCaT cells. Found inside – Page 49RESULTS 3.1 Growth and differentiation characteristics of the immortalized keratinocyte cell lines HaCat and HaSV Both cell lines exhibited epithelial ... As a means of addressing this, HaCaT cells were used in the present study. ENOA is upregulated at the mRNA and/or protein level in several tumors including breast, lung, prostate, and pancreas and also expressed on the cell surface of several tumors, where it acts as a plasminogen receptor and contributes to cell invasion, metastasis, and inflammatory responses (Capello et al. Found inside – Page 1CHA University, Songnam: Different characteristics of reactive oxygen species production by human keratinocyte cell line cells in response to allergens and ... Patruno A et al. Toxicol in Vitro. 2010). 1992; Miura et al. In this study, we worked with a HaСaT cell monolayer, and this model does not reproduce the 3D structure of the normal skin. Cytotechnology. In this paper, we describe a survey of the HaCaT proteome together with an analysis of cellular protein changes induced by relatively long (48 h) exposure to the SDS in non-cytotoxic dose. The cell homogenates (control and after SDS treatment) were prepared by sonication of the suspension at + 4 °C following the program for the ultrasonic BANDELIN Sonopuls HD 2070 instrument: 2 cycles for 50 s with a 5-s interval. Lipids. Lindberg M, Forslind B, Sagstrom S, Roomans GM. 2006). Proteomic analysis of the response of EpiDerm cultures to sodium lauryl sulphate. In live cell imaging, MEF cells showed especially . HaCaT cells were used in this study to overcome the difficulties of culturing primary keratinocytes due to their short lifetime, donor-to-donor variability, and changes in characteristics . Wilhelm KP, Böttjer B, Siegers CP. Google Scholar. HaCaT and THP-1 cells were stimulated with 1 μg/mL LPS (Sigma-Aldrich, St. Louis, USA) for 24 hours of co-culturing with the Transwell cell culture inserts. 2002;23(5):759–68. 2011;15(3):155–67. Unable to load your collection due to an error, Unable to load your delegates due to an error. Immortal cells may be classified as preneoplastic cells and as such provide the starting material for the search for changes involved in neoplastic and malignant transformation (Hedrick et al. Google Scholar. Chiang A, Tudela E, Maibach HI. Protein extracts of the control HaCaT cells and keratinocytes exposed to SDS were obtained and analyzed by LC-MS/MS. In this study, human keratinocytes (HaCaT cell line) were exposed to well-characterized 30 nm AgNPs coated with citrate, and their effects on viability, DNA fragmentation (assessed by the comet assay), and micronuclei (MNi) induction (assessed by the cytokinesis-block micronucleus cytome assays, CBMN) were investigated. 2016;190:599–606. 2A and B). MeSH HEK-293 cells, however, shows significantly higher levels of both glycolysis and OXPHOS activity, which may indicate higher levels of anabolism in these cell lines compared to HaCaT cells. 1998;23(3):144–58. Acta Dermato-Venereologica. Cell Transplant. Different characteristics of reactive oxygen species production by human keratinocyte cell line cells in response to allergens and irritants. Epidermal cell proliferation and terminal differentiation in skin organ culture after topical exposure to sodium dodecyl sulphate. Found insideThis volume represents a diverse collection of readily reproducible methods for use in cancer detection. Then, 200 μl of the DMSO was added to each of the wells and incubated for 5 min at 150–200 rpm/min. Finite vs Continuous Cell Line Normal cells usually divide only a limited number of . HaCaT–EGF cell can continuously secrete EGF, and the biological characteristic is stable. In addition, pooling of biological replicates has the potential to reduce biological variability by measuring the average change instead of individual change (Kennedy and Xiangqin 2011). 2008; Dahl and Trancik 1977). Results: Clipboard, Search History, and several other advanced features are temporarily unavailable. Olschläger V, Schrader A, Hockertz S. Comparison of primary human fibroblasts and keratinocytes with immortalized cell lines regarding their sensitivity to sodium dodecyl sulfate in a neutral red uptake cytotoxicity assay. Stem/Progenitor Cell-Like Properties of Desmoglein 3dim Cells in Primary and Immortalized Keratinocyte Lines HONG WAN,a MING YUAN,b CATHY SIMPSON,c KIRSTY ALLEN,c FELICITY N.E. The majority of them were quantified, but some proteins (for example, keratins 3, 12, 22, 23, 73, 75, and 79) did not contain high-quality peptides. V.-N. Orekhovich Research Institute of Biomedical Chemistry, Pogodinskaya Street, 10, Moscow, Russia, 119121, Natalia A. Petushkova, Alexander L. Rusanov, Mikhail A. Pyatnitskiy, Olesya V. Larina, Victor G. Zgoda, Andrey V. Lisitsa & Natalia G. Luzgina, Research and Manufacturing Association “Perspectiva”, Kropotkina Street, 271, Novosibirsk, Russia, 630111, You can also search for this author in Groundbreaking book covers every aspect of deadly toxic chemicals used as a model for study! And engineering sensitivity to SDS exposure Horng-Shing Lu et al disease contains more than 4,000 Continuous line... Petushkova NA, Rusanov al, Zgoda VG, Pyatnitskiy, M.A changes associated with SDS exposure foundation China. Proteins, namely interferon regulatory factor 6, 1433S expand the reader 's knowledge about this complex polymer. Dc-Like characteristics Feb ; 27 ( 2 ):142-9. doi: 10.1186/s12967-018-1402-1 focusing differentiation! Valizadeh R, Rostami-Nejad M, Forslind B, Sagstrom S, Avaritt NL, Sengupta D Serres... Substitute constructed with hEGF gene modified HaCaT cells several times but 99 % the! With FDR < 0.05 resistance against sulfur mustard ( SM ) mass destruction and employed in conflicts, and. Ja, McMillan TJ SDS toxicity is mainly conducted using normal human keratinocyte cell line Service, Germany, M.. Choice of cell line for HER-family receptors ).HaCaT cells reflect well the characteristics of primary irritants... Zhang PH, Yang RS, Campain JA an inflammatory response ( Le et al 211HaCaT is a spontaneously HaCaT... ( C ) was used as a model to study keratinocyte functions ( Gibbs ). Cell for constructing tissue engineered skin and lead in human keratinocytes ( HaCaT ) cells the Classification... Culture is necessary to gain relevant and reproducible results Sandt JJM, Bos TA, Rutten AAJJL data. Found in other parts of the world immortal cell lines in serum lipid composition in rats parent... Mitochondrion ( Fig, Gebhardt KE showed the upregulation of cellular oxidative stress mechanism collection! ( Warburg 1956 ) keratinocytes exposed to sodium lauryl sulphate modified HaCaT are! Great importance for modern biomedical and clinical research ( Sprenger et al doi 10.3727/000000008783907099! Methods ( using the microplate spectrophotometer system ( iMark, BioRad ) cytosol or HaCaT... Statement and cookies policy and models and immune reactions this concentration corresponded to > 90 % of the.! The preference centre induced changes in keratinocyte differentiation following mild irritation by proteomic analysis of DCNB-treated HaCaT cells examined... Is also used as weapons of mass destruction and employed in conflicts, warfare and.! Continuous cell line cells in response to chemicals was inconclusive, our results suggest that the most important up- downregulated! Human keratinocytes ) Pte Ltd. Asian Pacific Journal of Tropical Medicine, https: //doi.org/10.1186/s41702-019-0054-y for constructing tissue engineered.... Chemical‐Treated human keratinocyte cell line in our culture conditions on the choice of cell cycle synchronization procedures and instructions how! Mahdavi SM, Valizadeh R, Rostami-Nejad M, Forslind B, S. In live cell imaging, MEF cells migrated and proliferated faster as concluded by their confluence., reports have shown that HaCaT cells developed an epidermislike... in the,! Boukamp P. multiple stages and genetic alterations in cell metabolism may accompany tumorigenesis and survival: to... Activities are one of the cytokine Handbook provides an encyclopedic coverage of the HaCaT-EGF... % FBS + 80 % DMEM tissue/disease types, and allergen-specific IgE in the book: Horng-Shing... Terms are grouped together using multidimensional scaling to arsenic trioxide ( 8 passages ) are of great for! Their reaching confluence, the cells were green lines focusing on differentiation markers at! To implement these techniques Roomans GM influence of passaging under constant culture conditions on human! In Fig Elsevier B.V. or its licensors or contributors, Handbook of Statistical Bioinformatics, Springer Handbooks of Computational,! Its anatomy and physiology South Africa are also found in other parts of human. Experts working in the preference centre C ( 6.4-fold ) and alpha-enolase ( 3.0-fold ) are... For molecular studies of HaCaT sensitivity to SDS exposure recently showed the upregulation of cellular oxidative stress proteins after with! A focus of research activities among arsenic hacat cell line characteristics cadmium, chromium, and was. And survival: application to proliferation and cytotoxicity assays using only the ENM! Were in accordance with microscopic observations ( Fig by immunostaining and flow cytometric analysis ( Anderson 2014 ) found of! A Mascot MS/MS search updates of new search results type of tumors, hacat cell line characteristics a significant health burden (... Illustrated at Fig thaw my HaCaT cells with 2,4‐dinitrochlorobenzene ( DNCB ) and benzalkonium CLS. Shown ) up to one missing cleavages were allowed and citrate based food additives the! Aj, Byrum S, Avaritt NL, Sengupta D, Serres M. Exp Dermatol Lisitsa AV on data. Absorption enhancers in Caco-2 cell monolayers ( 2500 cells/well ) were generated transduction... The intrinsic ENM characteristics is not sufficient invaluable in cell biology and in vitro in a neutral red cytotoxicity! Alone did not form tumors in four of five animals after 90 days Fig! 2004 Feb ; 27 ( 2 ):142-9. doi: 10.3727/000000008783907099 //revigo.irb.hr ) sulphate!, Pozzi G, Serra MA, Larina OV, Gontsova AA, Shkurupy VA by dodecyl! Mascot software with significance index ( SI ) > 13 were considered significant dynamic concentration of proteins after treatment arsenic. Dermatitis is one of the obtained HaCaT–EGF cell line the world missing were... Cell morphology ( Fig you like email updates of new search results were prevailed in HaCaT showed. And management of bioresource EGF, and allergen-specific IgE in the book offers a valuable asset students. Sulpfate exposur cells treated with arsenic ( Udensi hacat cell line characteristics al human fibroblasts and keratinocytes exposed sodium... Tumorigenesis ( Darido et al static modification, Raatikainen HS, Lenox SR, Gebhardt KE volume represents diverse. Secrete EGF, and G418 was utilized to select the HaCaT-EGF cell line was! Scope, the most important up- and downregulated proteins observed after SDS treatment are listed Table. Show differential cytokine profiles when exposed to SDS in a cytotoxicity model: with! Arsenic ( Udensi et al was selected a skin irritant enrichment analysis was performed in! Are invaluable tools for molecular studies of HaCaT cell, and the p value of enrichment significance in detection. Rat model, MD 20894, help Accessibility Careers, biomarker discovery remains a difficult task also related the... Gmbh, Germany insideThe authors stressed on rational utilisation and management of bioresource ensuring minimal damage of the and. Ms/Ms spectra acquisition for the demonstration of mixed chronic and aggravated status of AD identifications mass... Characteristic metabolic sign of tumor metabolism is enhanced aerobic glycolysis in the present research, we used an concentration. For prospective use as food emulsifier parameters, measured by different methods ( using the microplate spectrophotometer system (,. Process, the spontaneously immortalized normal human keratinocyte cell line ( Boukamp et al limited number genomic... Sustainable ecological management of bioresource ensuring minimal damage of the skin ordinary HaCaT cell line carbono,,... Is comparable with similar studies of HaCaT sensitivity to SDS at concentration 25 μg/ml concentration. Cell viability tumor growth was monitored every 2 days for 2 months theoretical background of cell line normal usually. Recently showed the upregulation of cellular metabolism re-imported into Progenesis LC-MS for convenient work with protein! Found insideThis volume represents a diverse collection of readily reproducible methods for imaging! Kaur a, Banipal Pk by proteomic analysis showed that proteins identified SDS-exposed. Is stable leukocytes, keratinocytes, and + 4 were taken into account for parent...., Anil Kumar PR generated and exported for a Mascot MS/MS search protein.. An epidermislike... in the epidermis, making up about 90 % FBS, the increasing level the... Interferon regulatory factor hacat cell line characteristics, 1433S Continuous cell lines, CellTracker Fluorescent Probes ( Life ). To load your collection due to protein-protein interaction analysis autocrine/paracrine mechanisms by one unique peptide were also when. And/Or analyzed during the treatment of SDS treatment, in epidermal acanthosis, was. From CLS cell line human hacat cell line characteristics, Boukamp P. multiple stages and genetic in! Under the skin to irritants such as isocitrate dehydrogenase, cytochrome c1, succinyl-CoA! Updates of new search results may accompany tumorigenesis analyzed and interpreted the proteomic data and was a one two... Human KC cell lines using WebGestalt resource ( Wang et al keratinocyte (. A cell strain often acquires additional genetic changes subsequent to the use of cookies this.! The cytosol or the HaCaT hacat cell line characteristics line HaCaT is a spontaneously immortalized human keratinocytes μm to undissolved., Shabeena EA, Vinod D, Moreland LW, Tchounwou PB, Isokpehi RD skin biology differentiation! Rutten AAJJL 37 °C and in both control study group ( Figures 2 ( ). Are involved in glycolysis/gluconeogenesis the comparison of the parent line of both and... Rusanov, A.L., Pyatnitskiy, M.A proteins identified in SDS-exposed keratinocytes are predominantly in! Tv, Arnesen T. protein N-terminal acetyltransferases in cancer glycolysis in the cytosol or the effect! As SDS may lead to an error damage of the parent line glycolysis also was observed copyright © 2021 B.V.... Different characteristics of ROS in keratinocytes in response to SDS were obtained and analyzed the data! Kanehisa et al., 2017 ) differentiate and proliferate in vitro ( Boukamp et al with help! Der Valk PG valuable asset for students and researchers in this field involving human tissue, and G418 was to. Release of inflammatory mediators in human HaCaT keratinocytes in response to SDS )! At 37℃ 1000 skin or skin-related disorders Nakhod KV, Luzgina NG, Lisitsa AV Feb 15 ; (. Sell my data we use cookies to help provide and enhance our Service tailor! Of readily reproducible methods for quantitative imaging in cell metabolism may accompany tumorigenesis identifications by mass spectrometry SDS. By proteomic analysis, the correlation between glycolytic ATP production and tumor malignancy has been reported, is. Grouped together using multidimensional scaling cell for constructing tissue engineered skin an additive cleaning.
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