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首页> 美国卫生研究院文献>Journal of the Boston Society of Medical Sciences >Human skin in organ culture. Elaboration of proteolytic enzymes in the presence and absence of exogenous growth factors.
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Human skin in organ culture. Elaboration of proteolytic enzymes in the presence and absence of exogenous growth factors.

机译：人体器官文化中的皮肤。在存在和不存在外源性生长因子的情况下对蛋白水解酶进行精细化处理。

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Proteinase levels were assessed in organ culture fluids from human neonatal foreskin maintained under growth factor-free conditions and in the presence of a combination of growth factors (ie, epidermal growth factor, insulin, hydrocortisone, pituitary extract, and all-trans-retinoic acid). Analysis of culture fluids by gelatin zymography revealed the presence of 92-kd and 72-kd gelatinases. There was a greater amount of 92-kd gelatinase activity in the presence of growth factors whereas the levels of 72-kd gelatinase were similar in growth factor-free and growth factor-containing media. Experiments with keratinocytes and fibroblasts in monolayer culture and with isolated dermal tissue in organ culture indicated that the epithelial component was responsible for most of the 92-kd gelatinase activity whereas fibroblasts were primarily responsible for the 72-kd gelatinase activity. Activation with aminophenyl mercuric acetate, requirement for divalent cations, inhibition with EDTA, and insensitivity to inhibition with phenylmethyl sulfonyl fluoride indicated that both gelatinases were metalloproteinases. In additional studies, culture fluids were examined for the presence of plasminogen activator activity. This was detected in culture fluids from tissues maintained under both conditions but was increased in the growth factor-containing medium. The increased amount seen in the growth factor-containing medium appeared to be due almost entirely to a single factor, ie, all-trans-retinoic acid. In monolayer culture, both keratinocytes and fibroblasts produced plasminogen activator; the level was higher in keratinocyte culture fluids than in culture fluids from fibroblasts.
机译：在无生长因子的条件下，在存在多种生长因子（即表皮生长因子，胰岛素，氢化可的松，垂体提取物和全反式维甲酸）的情况下，对来自人类新生儿包皮的器官培养液中的蛋白酶水平进行了评估。 ）。用明胶酶谱法分析培养液表明存在92 kd和72 kd明胶酶。在存在生长因子的情况下，存在更多的92 kd明胶酶活性，而在无生长因子和含生长因子的培养基中，72 kd明胶酶的水平相似。单层培养中的角质形成细胞和成纤维细胞以及器官培养中的离体真皮组织的实验表明，上皮成分是大多数92 kd明胶酶活性的原因，而成纤维细胞主要是72 kd明胶酶活性的原因。氨基苯乙酸汞的活化，对二价阳离子的要求，EDTA的抑制作用以及对苯甲基磺酰氟的抑制作用不敏感表明这两种明胶酶都是金属蛋白酶。在其他研究中，检查了培养液中是否存在纤溶酶原激活剂活性。在两种条件下维持的组织的培养液中都检测到了这种现象，但是在含有生长因子的培养基中却增加了。在含有生长因子的培养基中看到的增加的量似乎几乎完全归因于单一因素，即全反式维甲酸。在单层培养中，角质形成细胞和成纤维细胞均产生纤溶酶原激活剂。角质形成细胞培养液中的水平高于成纤维细胞培养液中的水平。

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