Swartzendruber D C, Kitko D J, Wertz P W, Madison K C, Downing D T
Department of Dermatology, University of Iowa College of Medicine, IA 52242.
Arch Dermatol Res. 1988;280(7):424-9. doi: 10.1007/BF00429982.
Sheets of porcine stratum corneum were dispersed into individual corneocytes after 4 h in a solution consisting of 8 mM N,N-dimethyldodecylamine oxide and 2 mM sodium dodecylsulfate in phosphate-buffered isotonic saline, at 45 degrees C. With continued detergent treatment and moderate sonication, most of the cells lost their keratin contents and were then separated from the remaining intact cells by centrifugation in cesium chloride solution of density 1.280. Electron microscopy showed that the cell envelopes retained both the crosslinked protein envelope and its attached lipid envelope. The dry weight of envelopes was approximately 7% of the estimated dry weight of the original stratum corneum, while the corneocytes surviving intact also amounted to 7% of the starting weight. Mild alkaline hydrolysis of the corneocyte envelopes allowed the extraction of hydroxyceramides amounting to 10% of the dry weight of the envelopes. The procedure therefore provides isolated corneocyte envelopes suitable for studying both the protein and lipid components of this compound sheath.
猪角质层薄片在含8 mM N,N - 二甲基十二烷基氧化胺和2 mM十二烷基硫酸钠的磷酸盐缓冲等渗盐溶液中，于45摄氏度处理4小时后分散成单个角质形成细胞。随着去污剂处理的持续进行以及适度超声处理，大多数细胞失去了角蛋白成分，然后通过在密度为1.280的氯化铯溶液中离心，与其余完整细胞分离。电子显微镜显示，细胞包膜保留了交联的蛋白质包膜及其附着的脂质包膜。包膜的干重约为原始角质层估计干重的7%，而完整存活的角质形成细胞也占起始重量的7%。对角质形成细胞包膜进行温和的碱性水解可提取出占包膜干重10%的羟基神经酰胺。因此，该方法提供了适合研究这种复合鞘的蛋白质和脂质成分的分离角质形成细胞包膜。
