Weterings P J, Vermorken A J, Bloemendal H
Mol Biol Rep. 1980 Oct 31;6(3):153-8. doi: 10.1007/BF00775409.
A new technique has been used for culturing human keratinocytes. The cells grow on the basement membrane-like capsules of bovine lenses. Lens cells were removed from the capsules by rigid trypsinization. In order to exclude any contamination with remaining living cells the isolated capsules were irradiated with X-rays at a dose of 10,000 rad. In this way human epithelial cells can be brought in culture from individual hair follicles. Since feeder cells are not used in this culture technique, the biosynthesis of keratinocyte proteins can be studied in these cultures. The newly synthesized proteins can be separated into a water-soluble, a urea-soluble, and a urea-insoluble fraction. Product analysis has been performed on the first two fractions revealing protein patterns identical to those of intact hair follicles. Product analysis of the urea-soluble fractions of microdissected hair follicles shows that the protein pattern of the cultured keratinocytes resembles the protein pattern of the hair follicle sheath. Studies on the metabolism of benzo(a)pyrene revealed that the enzyme aryl hydrocarbon hydroxylase (AHH) is present in cultured hair follicle cells. A possible use of our culture system for eventual detection of inherited predisposition for smoking-dependent lung cancer is discussed.
一种新技术已被用于培养人角质形成细胞。这些细胞在牛晶状体的基底膜样胶囊上生长。通过强力胰蛋白酶消化从胶囊中去除晶状体细胞。为了排除残留活细胞的任何污染,将分离出的胶囊用10000拉德的X射线照射。通过这种方式,可以从单个毛囊中培养人上皮细胞。由于在这种培养技术中不使用饲养细胞,因此可以在这些培养物中研究角质形成细胞蛋白质的生物合成。新合成的蛋白质可以分离为水溶性、尿素溶性和尿素不溶性部分。对前两个部分进行了产物分析,结果显示其蛋白质模式与完整毛囊的蛋白质模式相同。对显微切割毛囊的尿素溶性部分进行的产物分析表明,培养的角质形成细胞的蛋白质模式类似于毛囊鞘的蛋白质模式。对苯并(a)芘代谢的研究表明,培养的毛囊细胞中存在芳烃羟化酶(AHH)。讨论了我们的培养系统最终用于检测吸烟依赖性肺癌遗传易感性的一种可能用途。