Measuring the Vital Function of the Skin Barrier
Session Number
MEDH 06
Advisor(s)
Dr. Bethany Perez White, Northwestern University, Feinberg School of Medicine
Discipline
Medical and Health Sciences
Start Date
17-4-2024 10:25 AM
End Date
17-4-2024 10:40 AM
Abstract
The epidermal compartment of skin balances proliferation for tissue self-renewal and terminal differentiation for essential barrier function. This study examines how keratinocytes, the cells that comprise the epidermis, build a barrier as they become terminally differentiated. To assess the function of the epidermal barrier, we used an epithelial voltohmmeter (EVOM2) to measure the transepithelial electrical resistance (TEER, Ω⋅cm2) of keratinocyte cultures, where increased resistance indicates a stronger barrier. Keratinocytes were isolated from human skin samples and cultured in M154 media specific for keratinocyte propagation. We directly compared the barrier function of proliferating versus differentiating keratinocytes over the course of seven days. Proliferating conditions were maintained by keeping cultures in low calcium (0.07 mM); differentiation was induced by increasing calcium levels to 1.2 mM (high calcium). We probed each well at three locations within the culture to ensure accurate measurements. Cells were plated at a density of 1.5E6 cells/well (well surface area: 4.67 cm2). We performed TEER over a course of seven days, taking measurements at different time points. We found that cells in a high calcium environment had statistically significantly higher resistance compared to those in low calcium. The results of this assay can help identify what perturbations may affect the barrier positively or negatively.
Measuring the Vital Function of the Skin Barrier
The epidermal compartment of skin balances proliferation for tissue self-renewal and terminal differentiation for essential barrier function. This study examines how keratinocytes, the cells that comprise the epidermis, build a barrier as they become terminally differentiated. To assess the function of the epidermal barrier, we used an epithelial voltohmmeter (EVOM2) to measure the transepithelial electrical resistance (TEER, Ω⋅cm2) of keratinocyte cultures, where increased resistance indicates a stronger barrier. Keratinocytes were isolated from human skin samples and cultured in M154 media specific for keratinocyte propagation. We directly compared the barrier function of proliferating versus differentiating keratinocytes over the course of seven days. Proliferating conditions were maintained by keeping cultures in low calcium (0.07 mM); differentiation was induced by increasing calcium levels to 1.2 mM (high calcium). We probed each well at three locations within the culture to ensure accurate measurements. Cells were plated at a density of 1.5E6 cells/well (well surface area: 4.67 cm2). We performed TEER over a course of seven days, taking measurements at different time points. We found that cells in a high calcium environment had statistically significantly higher resistance compared to those in low calcium. The results of this assay can help identify what perturbations may affect the barrier positively or negatively.