Effect of Lytic Enzymes Upon the Responsivity of Rat Adrenals<i>in Vitro</i>. I. Effect of Trypsin Upon the Steroidogenic Action of Reduced Triphosphopyridine Nucleotide

Halkerston, I. D. K.; Feinstein, Maurice B.; Hechter, Oscar

doi:10.1210/endo-83-1-61

Cited by 14 publications

(11 citation statements)

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“…The monolayer culture system with intact viable cells has considerable advantages in the study of the structure and function of endocrine cells. Tissue slices and similar preparations can only be maintained in vitro for limited periods of time, and may show abnormal responses due to the presence of damaged cells (Halkerston, Feinstein & Hechter, 1968). Damaged cells may also be present in fresh cell suspensions, and organ cultures are often subject to considerable degeneration.…”

Section: Discussionmentioning

confidence: 99%

Morphological Responses to Corticotrophin and Cyclic Amp by Adult Rat Adrenocortical Cells in Monolayer Culture

O'Hare¹,

Am²

1973

Journal of Endocrinology

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Confluent monolayer cultures of cells from the zona fasciculata andzona reticularis of the normal adult rat adrenal cortex were maintained with or without corticotrophin (ACTH) for up to 4 months, without proliferation of adrenal cells. Proliferating fibroblast-like cells, however, eventually overgrew the adrenal monolayer in cultures both with and without ACTH. Adrenocortical cells in culture, maintained without ACTH, spread rapidly to form a confluent monolayer, whereas cell spreading was markedly inhibited in the presence of ACTH. Exposure of previously unstimulated cells to ACTH or cyclic AMP caused the adrenal cells to retract with loss of confluence, the process being reversed when ACTH or cyclic AMP was withdrawn. Ultrastructural features of cells cultured with ACTH were typical of normal adrenocortical cells; in cultures without ACTH they were similar to those of adrenocortical cells found in the hypophysectomized rat.

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Section: Discussionmentioning

confidence: 99%

Morphological Responses to Corticotrophin and Cyclic Amp by Adult Rat Adrenocortical Cells in Monolayer Culture

O'Hare¹,

Am²

1973

Journal of Endocrinology

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“…A number of successful attempts have been made recently to produce viable single cell suspensions from a variety of tissues (e .g ., liver [32,33], thyroid [34], adrenal [5,35,36], corpus luteum [37], and exocrine pancreas [38]), including the anterior pituitary (2-6, [39][40][41] . Most of these procedures have involved the use of collagenase, hyaluronidase, trypsin, pronase, EDTA, THE JOURNAL OF CELL BIOLOGY .…”

Section: Cell Dissociation Proceduresmentioning

confidence: 99%

Hormone Secretion by Cells Dissociated From Rat Anterior Pituitaries

Hopkins

Farquhar

1973

The Journal of Cell Biology

240

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A new procedure has been developed for dissociating anterior pituitary tissue and producing a viable suspension of single cells . The procedure involves incubation of small tissue blocks in 1 mg/ml trypsin (15 min), followed by incubation in 8µg/ml neuraminidase and 1 mM EDTA (15 min), followed by mechanical dispersion . Cell yields are -55%, based on recovered DNA. By electron microscopy five types of secretory cells (somatotrophs, mammotrophs, thyrotrophs, gonadotrophs, and corticotrophs) plus endothelial and follicular cells can be identified and are morphologically well preserved up to 20 h after dissociation . Throughout this period, the cells incorporate linearly [ 3 H]leucine into protein for up to 4 h at a rate 90% greater than hemipituitaries, and they synthesize, transport intracellularly, and release the two major pituitary secretory products, growth hormone and prolactin . Immediately after dissociation the cells' ability to respond to secretogogues (high K+ and dibutyryl cyclic AMP) is impaired, but after a 6-12-h culture period, the cells apparently recover and discharge 24% and 52%, respectively, of their content of prelabeled growth hormone over a 3-h period in response to these two secretogogues . This represents a stimulation of 109% and 470% over that released by cells incubated in control medium . The results demonstrate that function and morphologic integrity are preserved in this cell system . Therefore it is suitable for the study of various aspects of pituitary secretion and its control .

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“…For the dispersion of adrenal cells, several methods have been developed. Kloppenborg et al(1968) and Haning et al(1970) used collagenase, and Halkerston and Feinstein (1968) employed a combined use of five enzymes. Swallow and Sayers (1969), and Sayers et al(1971) reported a unique method using trypsin as a dispersing enzyme.…”

mentioning

confidence: 99%

A Simple Method for the Preparation of ACTH Responsive Isolated Cells from Rat Adrenals

Nakamura

1971

Endocrinol Japon

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Effect of Lytic Enzymes Upon the Responsivity of Rat Adrenalsin Vitro. I. Effect of Trypsin Upon the Steroidogenic Action of Reduced Triphosphopyridine Nucleotide

Cited by 14 publications

References 0 publications

Morphological Responses to Corticotrophin and Cyclic Amp by Adult Rat Adrenocortical Cells in Monolayer Culture

Morphological Responses to Corticotrophin and Cyclic Amp by Adult Rat Adrenocortical Cells in Monolayer Culture

Hormone Secretion by Cells Dissociated From Rat Anterior Pituitaries

A Simple Method for the Preparation of ACTH Responsive Isolated Cells from Rat Adrenals

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