HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes.
HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes.
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DOI：
10.1155/2017/7435621
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发表时间：
2017
影响因子：
4.6
通讯作者：
Dell'Agli M
中科院分区：
文献类型：
--
作者：
Colombo I;Sangiovanni E;Maggio R;Mattozzi C;Zava S;Corbett Y;Fumagalli M;Carlino C;Corsetto PA;Scaccabarozzi D;Calvieri S;Gismondi A;Taramelli D;Dell'Agli M
培养的原代人角质形成细胞经常用于免疫和炎症反应的研究；然而，对实验数据的解释可能会因为供体之间的差异、相对较短的培养寿命和传代之间的差异而变得复杂。为了规范角质形成细胞的体外研究，我们研究了HaCaT细胞的使用，HaCaT细胞是一种长寿命，自发永生化的人角质形成细胞系，能够在体外分化，作为一种合适的模型，跟踪炎症和修复介质在TNFα或IL-1β反应中的释放。考虑了不同的处理条件（血清存在或不存在）和分化刺激（细胞密度随培养时间的增加和细胞外钙的升高）。采用ELISA和Multiplex检测技术监测细胞因子和趋化因子的产生。综上所述，研究结果表明，培养基中的Ca2+浓度、细胞密度和血清的存在在不同程度上影响HaCaT细胞释放促炎介质。此外，HaCaT细胞在低Ca2+培养基中维持80%的融合，在细胞因子产生方面与正常角质形成细胞相似，这表明HaCaT细胞可能是研究抗炎干预/治疗皮肤病的有用模型。
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.
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影响因子：
3
作者：
Pfundt, R;Wingens, M;Schalkwijk, J
通讯作者：
Schalkwijk, J
影响因子：
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Dimoudis, N
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