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Primary tissue culture of human adult adrenocortical cells. Methodology and electron microscopic observations on ACTH-deprived and ACTH-treated cortical cells

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Summary

A method of primary tissue culture involving both disaggregation of cells by repeated exposure of small tissue fragments to a solution of trypsin, collagenase and hyaluronidase and explantation of the residual tissue fragments intermingled with isolated cells onto polyethylene discs, has been shown to be adequate for the prolonged maintenance (up to 30 days) in vitro of cells arising from decapsulated adult human adrenocortical tissue. The technique and its critical points are discussed.

Adrenocortical cells were organized both as outgrowing columns from microexplants or as variously sized islets of monolayered cells. The ultrastructural features of ACTH-deprived adrenocortical cells (i.e., mitochondria with laminar cristae, endoplasmic reticulum mainly consisting of rough profiles, abundance of lipid droplets and β-glycogen particles) suggest that the cells dedifferentiate and retain practically no steroidogenic activity.

After 2 days of ACTH-treatment, cultured parenchymal cells were found to be quite similar to the zona fasciculata elements of the normal human adrenal cortex. They were grouped in islets of about 50–100 cells. Rough endoplasmic reticulum had decreased, but smooth endoplasmic reticulum showed focal proliferation. The pleomorphic mitochondria with laminar cristae, transformed into a homogeneous population of round or ovoid mitochondria containing tubulo-vesicular cristae. Lipid droplets and glycogen particles were decreased in number.

After 7 days of daily treatment with ACTH, the cortical elements, whose nucleus and cytoplasm seemed to be enlarged, were arranged in clusters formed by up to 300 monolayered elements, in which dividing cells were consistently observed. Their cytoplasm was filled with a meshwork of smooth reticulum tubules, in which scantly ribosome-studded profiles and occasional small stacks of granular cisternae were embedded. Mitochondria were similar to those of the 2 days ACTH-treated cultures. Lipid droplets and glycogen particles were absent. The functional significance of these structural changes as well as the possible mechanism underlying the differentiative effect of ACTH are discussed.

Primary cultures of human adult adrenals are proposed as a new tool for studies into the physiopathology of the adrenocortical cells under carefully controlled experimental conditions.

mis|It is a pleasure to acknowledge our thanks to Drs. F. Mantero and C. Eccher for kindly supplying the normal human adrenocortical tissue. Thanks are also due to Mr. G. Gottardo for his excellent technical assistance.

mis|This work was partly supported by a contract with the CNR-Italy (C.T. 73.00663.04).

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Authors and Affiliations

  1. Department of Anatomy, Laboratories of Tissue Culture and Electron Microscopy, University of Padua, Padua, Italy

    Ubaldo Armato,  Gastone G. Nussdorfer, Paola G. Andreis, Giuseppina Mazzochi & Enrica Draghi

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  1. Ubaldo Armato
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  2. Gastone G. Nussdorfer
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  3. Paola G. Andreis
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  4. Giuseppina Mazzochi
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  5. Enrica Draghi
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A preliminary report on part of this work was given at the Annual Meeting of the European Tissue Culture Society, Genua, May 1974, and at the Annual Meeting of the Société Française de Microscopie Electronique, Rennes, May 1974, and published as short communication in The Journal of Endocrinology 63, 247, 1974.

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Armato, U., Nussdorfer, G.G., Andreis, P.G. et al. Primary tissue culture of human adult adrenocortical cells. Methodology and electron microscopic observations on ACTH-deprived and ACTH-treated cortical cells. Cell Tissue Res. 155, 155–180 (1974). https://doi.org/10.1007/BF00221351

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