Yokozeki H, Saga K, Sato F, Sato K
Marshall Dermatology Research Laboratories, Department of Dermatology, University of Iowa College of Medicine, Iowa City 52242.
Am J Physiol. 1990 Jun;258(6 Pt 2):R1355-62. doi: 10.1152/ajpregu.1990.258.6.R1355.
Methacholine (MCh)- and isoproterenol (Iso)-stimulated 14CO2 production was compared between freshly dissociated rhesus sweat secretory coil cells (mainly clear cells) and cultured cells (grown on a collagen-coated plastic plate) derived from native cells. 14CO2 production was enhanced by MCh and by Iso in native coil cells (but not in ductal cells) in a pharmacologically specific and dose-dependent manner. 14CO2 production in subcultured coil cells (19-45 days in culture) was only one-third to one-fifth that of native cells. MCh-stimulated 14CO2 production was inhibited by ouabain and furosemide in both native and cultured coil cells. A decrease in 14CO2 production, of about one-half, was already evident in primary cells cultured for less than 1 wk. The decreased pharmacological responsiveness of the cultured coil cells was seen, although the cultured cells showed the typical epithelioid appearance, abundant mitochondria, the occasional presence of intercellular lacunae resembling intercellular canaliculi, and the persistence of immunoreactive keratin. We conclude that 1) a primary culture of sweat gland cells can be initiated from dissociated cells; 2) cultured sweat secretory coil cells qualitatively, but not quantitatively, retain the pharmacological responsiveness and transport activity of the native cells as determined by 14CO2 production; 3) collagenase-dissociated cells represent an excellent in vitro system for the study of glandular function at the cellular level; and 4) the decrease in pharmacological responsiveness is not simply due to trypsin treatment during harvesting of cultured cells, because that of organ-cultured, intact, secretory coils also declines with time of culture.
比较了新鲜解离的恒河猴汗腺分泌盘曲部细胞（主要是透明细胞）与源自天然细胞的培养细胞（生长在胶原包被的塑料板上）对乙酰甲胆碱（MCh）和异丙肾上腺素（Iso）刺激的(^{14}CO_2)生成情况。在天然盘曲部细胞（而非导管细胞）中，MCh和Iso以药理学特异性和剂量依赖性方式增强了(^{14}CO_2)的生成。传代培养的盘曲部细胞（培养19 - 45天）中的(^{14}CO_2)生成量仅为天然细胞的三分之一至五分之一。在天然和培养的盘曲部细胞中，哇巴因和呋塞米均抑制MCh刺激的(^{14}CO_2)生成。在培养不到1周的原代细胞中，(^{14}CO_2)生成量已明显下降约一半。尽管培养细胞呈现典型的上皮样外观、有丰富的线粒体、偶尔存在类似细胞间小管的细胞间腔隙且持续存在免疫反应性角蛋白，但仍观察到培养的盘曲部细胞的药理学反应性降低。我们得出以下结论：1）可以从解离的细胞开始进行汗腺细胞的原代培养；2）通过(^{14}CO_2)生成量测定，培养的汗腺分泌盘曲部细胞在质量上而非数量上保留了天然细胞的药理学反应性和转运活性；3）胶原酶解离的细胞代表了用于在细胞水平研究腺体功能的优秀体外系统；4）药理学反应性的降低并非仅仅由于培养细胞收获过程中的胰蛋白酶处理，因为器官培养的完整分泌盘曲部的反应性也随培养时间而下降。
