HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes
Cultured primary human keratinocytes are frequently employed for studies of immunological and inflammatory responses; however, interpretation of experimental data may be complicated by donor to donor variability, the relatively short culture lifetime, and variations between passages. To standardize...
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| Format: | Article |
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Wiley
2017-01-01
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| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/2017/7435621 |
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| author | Irma Colombo Enrico Sangiovanni Roberta Maggio Carlo Mattozzi Stefania Zava Yolanda Corbett Marco Fumagalli Claudia Carlino Paola Antonia Corsetto Diletta Scaccabarozzi Stefano Calvieri Angela Gismondi Donatella Taramelli Mario Dell’Agli |
| author_facet | Irma Colombo Enrico Sangiovanni Roberta Maggio Carlo Mattozzi Stefania Zava Yolanda Corbett Marco Fumagalli Claudia Carlino Paola Antonia Corsetto Diletta Scaccabarozzi Stefano Calvieri Angela Gismondi Donatella Taramelli Mario Dell’Agli |
| author_sort | Irma Colombo |
| collection | DOAJ |
| description | Cultured primary human keratinocytes are frequently employed for studies of immunological and inflammatory responses; however, interpretation of experimental data may be complicated by donor to donor variability, the relatively short culture lifetime, and variations between passages. To standardize the in vitro studies on keratinocytes, we investigated the use of HaCaT cells, a long-lived, spontaneously immortalized human keratinocyte line which is able to differentiate in vitro, as a suitable model to follow the release of inflammatory and repair mediators in response to TNFα or IL-1β. Different treatment conditions (presence or absence of serum) and differentiation stimuli (increase in cell density as a function of time in culture and elevation of extracellular calcium) were considered. ELISA and Multiplex measurement technologies were used to monitor the production of cytokines and chemokines. Taken together, the results highlight that Ca2+ concentration in the medium, cell density, and presence of serum influences at different levels the release of proinflammatory mediators by HaCaT cells. Moreover, HaCaT cells maintained in low Ca2+ medium and 80% confluent are similar to normal keratinocytes in terms of cytokine production suggesting that HaCaT cells may be a useful model to investigate anti-inflammatory interventions/therapies on skin diseases. |
| format | Article |
| id | doaj-art-724085f532e0407d8be59433475e4abb |
| institution | Kabale University |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 2017-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-724085f532e0407d8be59433475e4abb2025-02-03T01:09:39ZengWileyMediators of Inflammation0962-93511466-18612017-01-01201710.1155/2017/74356217435621HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human KeratinocytesIrma Colombo0Enrico Sangiovanni1Roberta Maggio2Carlo Mattozzi3Stefania Zava4Yolanda Corbett5Marco Fumagalli6Claudia Carlino7Paola Antonia Corsetto8Diletta Scaccabarozzi9Stefano Calvieri10Angela Gismondi11Donatella Taramelli12Mario Dell’Agli13Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano, ItalyDipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano, ItalyDipartimento di Medicina Sperimentale, Università di Roma La Sapienza, Viale Regina Elena 324, 00161 Roma, ItalyDipartimento di Medicina Interna e Specialità Mediche, UOC di Clinica Dermatologica, Università di Roma “Sapienza”, Viale del Policlinico 155, 00161 Roma, ItalyDipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano, ItalyDipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano, ItalyDipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano, ItalyCenter for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, Viale Regina Elena 291, 00161 Roma, ItalyDipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano, ItalyDipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano, ItalyDipartimento di Medicina Interna e Specialità Mediche, UOC di Clinica Dermatologica, Università di Roma “Sapienza”, Viale del Policlinico 155, 00161 Roma, ItalyDipartimento di Medicina Molecolare, Università di Roma La Sapienza, Viale Regina Elena 291, 00161 Roma, ItalyDipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano, ItalyDipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Via Balzaretti 9, 20133 Milano, ItalyCultured primary human keratinocytes are frequently employed for studies of immunological and inflammatory responses; however, interpretation of experimental data may be complicated by donor to donor variability, the relatively short culture lifetime, and variations between passages. To standardize the in vitro studies on keratinocytes, we investigated the use of HaCaT cells, a long-lived, spontaneously immortalized human keratinocyte line which is able to differentiate in vitro, as a suitable model to follow the release of inflammatory and repair mediators in response to TNFα or IL-1β. Different treatment conditions (presence or absence of serum) and differentiation stimuli (increase in cell density as a function of time in culture and elevation of extracellular calcium) were considered. ELISA and Multiplex measurement technologies were used to monitor the production of cytokines and chemokines. Taken together, the results highlight that Ca2+ concentration in the medium, cell density, and presence of serum influences at different levels the release of proinflammatory mediators by HaCaT cells. Moreover, HaCaT cells maintained in low Ca2+ medium and 80% confluent are similar to normal keratinocytes in terms of cytokine production suggesting that HaCaT cells may be a useful model to investigate anti-inflammatory interventions/therapies on skin diseases.http://dx.doi.org/10.1155/2017/7435621 |
| spellingShingle | Irma Colombo Enrico Sangiovanni Roberta Maggio Carlo Mattozzi Stefania Zava Yolanda Corbett Marco Fumagalli Claudia Carlino Paola Antonia Corsetto Diletta Scaccabarozzi Stefano Calvieri Angela Gismondi Donatella Taramelli Mario Dell’Agli HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes Mediators of Inflammation |
| title | HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes |
| title_full | HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes |
| title_fullStr | HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes |
| title_full_unstemmed | HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes |
| title_short | HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes |
| title_sort | hacat cells as a reliable in vitro differentiation model to dissect the inflammatory repair response of human keratinocytes |
| url | http://dx.doi.org/10.1155/2017/7435621 |
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