Spatial quantification and classification of skin response following perturbation using organotypic skin cultures

Abstract

Integrating organotypic skin cultures, whole-slide scanning and subsequent image processing provides bioinformatics with a novel source of spatial expression data. We here used this approach to quantitatively describe the effect of treating organotypic skin cultures with sodium dodecyl sulphate in a non-corrosive concentration. We first measured the differentiation-related spatial expression gradient of Heat-Shock-Protein 27 in a time series of up to 24 h. Secondly, a multi-dimensional tissue classifier for predicting skin irritation was developed based on abstract features of these profiles. We obtained a high specificity of 0.94 and a sensitivity of 0.92 compared with manual classification. Our results demonstrate that the integration of tissue cultures, whole-slide scanning and image processing is well suited for both the standardized data acquisition for systems biological tissue models and a highly robust classification of tissue responses.

Stathmin Regulates Keratinocyte Proliferation and Migration during Cutaneous Regeneration

“…In order to exclude the possibility that the lack of Stathmin induction in the epidermal migration tongue was a specific phenomenon of murine wound healing, its expression was analyzed in an organotypic coculture (OTC) model consisting of primary human keratinocytes and fibroblasts after including 8 mm punch wounds [24]. Specimens were collected after different time points and processed for Stathmin staining.…”

“…For this, an automated and quantitative approach correlating Ki67 positivity (as measure for keratinocyte proliferation) and Stathmin staining was used [24]. …”

“…Epidermal keratinocytes used for EFT skin cultures were derived from human neonatal foreskin and were equilibrated and cultivated according to the manufacturers’ recommendation. EFT cultivation and specimen processing have been described elsewhere [24]. Samples were wounded using 8 mm circular biopsy punches and further cultured at 37°C in a 5% CO 2 incubator in EFT maintaining medium (MatTek, Ashland, MA, USA).…”

Risk factor-dependent dynamics of atopic dermatitis: modelling multi-scale regulation of epithelium homeostasis

“…For this purpose, it is essential to develop a new way to continuously measure the barrier integrity over several hours to a day; for example, by non-invasive measurement of transepidermal water loss and skin hydration measurements in AD patients [52] or by use of organotypic cultures, which allow temporal and spatial quantitative characterization of the skin [53]. Validation of our results through such studies will contribute to a more efficient, patient-specific treatment of AD and other atopic diseases, since the risk factor triggering the disease condition in a particular patient can be deduced from the time course of the barrier integrity.…”

“…An empirical validation of our model predictions, particularly the association between the four risk factors and dynamic model behaviours (figure 5), and implementing the prediction in dermatological practice will require an empirical system-level investigation of the epithelium to measure the barrier integrity and inflammation over time for different phenotypes and environmental conditions. For this purpose, it is essential to develop a new way to continuously measure the barrier integrity over several hours to a day; for example, by non-invasive measurement of transepidermal water loss and skin hydration measurements in AD patients [52] or by use of organotypic cultures, which allow temporal and spatial quantitative characterization of the skin [53]. Validation of our results through such studies will contribute to a more efficient, patient-specific treatment of AD and other atopic diseases, since the risk factor triggering the disease condition in a particular patient can be deduced from the time course of the barrier integrity.…”