血中ACTH様活性の定量的検定に関する研究

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概要

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• The author has tried to delvelop an accurate bioassay method of ACTH-like activity in human plasma, which can be used for daily routine clinical investigation, <BR>The details of this method and experimental background of procedures are reporeted in the following. <BR>Bioassay procedure <BR>1. Preparation of slices of adrenal gland <BR>Adrenal glands of 4 to 5 years old cows were removed and stored in ice-cold refrigerator immediately after slaughtering, and cut in thin slices with a razor blade within 45 minutes. <BR>Only left adrenal glands were used for convenience, and were firstly decapsulated and freed from marrow substances from the inside. <BR>It has been proved histologically that these slices consisted mainly of Z. fasciculata and Z. reticularis. <BR>Each slice was approximately 0.3 mm thin, weighed about 25 mg, and each slice of adrenal was kept in ice-cold normal saline separately. <BR>2. Minimization of individual variation <BR>Four slices from different animals (weighing about 100 mg) were put in one flask which contained 4 ml of KRBG (Krebs-Ringer bicarbonate glucose 200 mg%) as incubation medium. This procedure was necessary to minimize possible fluctuation of the response of individual slices to ACTH. <BR>3. Pre-incubation <BR>The first step of incubations was carried out for 60 minutes with a metabolic shaker at 37°C under gas phase of 95% O<SUB>2</SUB> and 5% CO<SUB>2</SUB>. <BR>At the end of incubation, the slices were blotted and washed 3 times with cooled normal saline. After this incubation, the biological conditions of slices were well standardized and the sensitivity of slices to ACTH was markedly enhanced thus this procedure was one of the imoprtant points of the assay. <BR>4. Final-incubation <BR>Final incubation was carried out for 90 minutes under the same conditions except for the incubation medium which contained the given amount of ACTH as standard or four times diluted plasma as unknown sample. <BR>5. Measurement of released corticosteroids

• 日本内分泌学会の論文

著者

• 安井 博和

  神戸医科大学第二内科学教室

関連論文

• 血中ACTH様活性の定量的検定に関する研究

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