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An Improved Technique for the Preparation of Isolated Rat Adrenal Cells: A Sensitive, Accurate and Specific Method for the Assay of ACTH
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1971
Year
Animal PhysiologyImproved TechniqueAdrenal GlandBiochemistryEndocrine MechanismMedicinePhysiologyBioanalysisMechanical AgitationSpecific MethodMetabolismTryptic DigestionAdrenal DiseaseClinical ChemistryEndocrinologyPharmacologyRat Adrenal TissueGlucocorticoid
Rat adrenal tissue is dispersed with a combination of tryptic digestion and mechanical agitation. Production of corticosterone by the isolated cells in response to a given quantity of ACTH is increased when the calcium concentration of the incubation medium is increased to 3 times that of KRB. Production of corticosterone is enhanced when BSA is added to the medium; production is greater at 0.5% than at 2.0 or 3.0% BSA. Minimum effective dose is 0.1 μU (1.0 pg) of ACTH. The accuracy of the isolated adrenal cell method for assay of ACTH is excellent; λ = 0.01–0.06 compared with λ =0.15–0.25 for other currently employed bioassay methods. In addition, the cells are highly selective; they do not respond to relatively large quantities (107 pg) of insulin, glucagon, oxytocin, vasopressin or angiotensin II. At 107 pg, α-MSH exhibits a weak action corresponding to that exhibited by 10 pg of ACTH. (Endocrinology88: 1063, 1971)