Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu, Tokyo 183-8509, Japan.
Res Vet Sci. 2020 Jun;130:48-51. doi: 10.1016/j.rvsc.2020.02.011. Epub 2020 Feb 19.
Ceramides (CERs) in the stratum corneum (SC) are known to play a crucial role in determining skin barrier function in dogs. We aimed to develop an in vitro culture system that synthesized epidermal CER classes to better understand the synthesis of CER classes in canine SC-. Canine keratinocyte cells (MSCEK) at appropriate confluency were incubated with high Ca (1.8 mM CaCl) supplemented serum-free medium. Eight days post Ca application, the surface of cultured MSCEK was broadly stained with anti-loricrin antibody implying that the keratinocytes had stratified into stratum granulosum. MSCEK cells synthesized heterogenous epidermal CERs, similar to those seen during the stratification of canine keratinocytes. CER fractions obtained from MSCEK cells were comparable to those from canine SC, including CER[EOS] (combination of ω-hydroxy fatty acids and sphingosines), CER[NP] (combination of non-hydroxy fatty acids and phytosphingosines), and CER[EOP] (combination of ω-hydroxy fatty acids and phytosphingosines), all of which are lowered in the SC during canine atopic dermatitis. Thus, the present study provides a simple culture system as a tool for in-depth analysis of CER production in canine keratinocytes.
角质层中的神经酰胺（CERs）在决定犬皮肤屏障功能方面起着至关重要的作用。本研究旨在开发一种体外培养体系，以更好地了解犬角质层中 CER 类的合成。将适当汇合的犬角质形成细胞（MSCEK）与高钙（1.8 mM CaCl）补充的无血清培养基孵育。钙处理 8 天后，用抗板层素抗体广泛染色培养的 MSCEK 表面，这表明角质形成细胞已分层到颗粒层。MSCEK 细胞合成了异质的表皮 CER，类似于犬角质形成细胞分层过程中观察到的 CER。从 MSCEK 细胞中获得的 CER 馏分与犬角质层中的 CER 相似，包括 CER[EOS]（ω-羟基脂肪酸和神经酰胺的组合）、CER[NP]（非羟基脂肪酸和植物鞘氨醇的组合）和 CER[EOP]（ω-羟基脂肪酸和植物鞘氨醇的组合），所有这些在犬特应性皮炎时角质层中含量降低。因此，本研究提供了一种简单的培养体系，可作为深入分析犬角质形成细胞 CER 产生的工具。
