Líf - og heilbrigðisvísindaráðstefna Háskóla Íslands 2023

Optimizing skin cell culture model to study epithelial barrier integrity

Soffía Rún Skúladóttir, Ari Jón Arason, Arna Steinunn Jónasardóttir, Bryndís Valdimarsdóttir, Thorarinn Gudjonsson

Introduction. Skin diseases are undervalued in the burden they cause the global population. Skin diseases are rarely fatal, and consequently attract less attention. Current treatments are largely focused on minimising symptoms such as itching and inflammation. Azithromycin (Azm) is an antibiotic that is commonly used to treat respiratory infections. We have previously shown that Azm enhances the integrity of the airway epithelium. The purpose of this project is to optimize the culture conditions of the keratinocyte cell line HaCaT with the aim of using this model to study skin epithelial barrier integrity. Methods. HaCaT was cultured in monolayer and air-liquid interface (ALI) condition. Analysis was done to compare different cell culture media DMEM+10% FBS, DMEM+3D, 3D and Keratinocyte growth medium (KGM). Epithelial integrity was evaluated by measuring transepithelial electrical resistance (TEER). Cell cultures were analysed by using immunostaining and RT-PCR. Results. Expression of skin markers (p63, keratin 1, keratin 14, transglutaminase-1 and filaggrin) in monolayer and ALI were most notable in KGM and DMEM+3D, respectively. In ALI, the highest TEER was achieved in 3D/DMEM medium. Histology analysis show that HaCaT cells capture many aspects of epidermis, including basal location of p63 positive cells and expression of involucrin and cytokeratins 1, 10 and 14. Furthermore, Azm treatment in ALI culture resulted in higher TEER, increased thickness of cellular layers and clear epidermal differentiation. Conclusions. HaCaT cells can capture functional and phenotypical architecture of the skin with prominent epithelial integrity which is further enhanced when cells are treated with Azm.


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