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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 244 (1986), S. 69-76 
    ISSN: 1432-0878
    Keywords: Adrenal medulla ; Tissue culture ; NGF ; Neuritic outgrowth ; Histofluorescence ; Cercopithecus aethiops ; Cebus apella
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Chromaffin cells from the monkey adrenal medulla were maintained in vitro in the presence of nerve growth factor (NGF) and the neuronal properties of these cells were assessed. Single-cell preparations were obtained by collagenase-trypsin treatment of the minced adrenal medulla tissue. Cells assumed a glandular to epithelioid morphology after twenty-four hours of culture. Twelve percent of these cells were shown to extend neurites spontaneously after five days. NGF-stimulated neuritic outgrowth from most cells after five days of culture and these neurites remained for at least three weeks. Cells exhibited intense histofluorescence for catecholamines even after three weeks in vitro in the presence of NGF and positive staining for tyrosine hydroxylase and dopamine beta hydroxylase could be detected by immunocytochemistry. Moreover, the chromaffin cells were shown to bind tetanus toxin, which is a specific marker for neurons. Tetanus toxin labelling was not dependent upon the presence of neurites on these cells. Transmission electron microscopy indicated that cultured cells contained numerous dense-core vesicles similar to noncultured medulla cells. Many of the neurites possessed the morphological features of axons; long varicose processes resembling noradrenergic fibers were identified by catecholamine histofluorescence and tyrosine hydroxylase immunocytochemistry. Microtubular arrays, in an axonal-like organization pattern, were seen ultrastructurally along with the presence of many dense-core vesicles. These data support the potential of adult primate chromaffin cells as a source of sympathetic neuronal tissue for neural transplantation.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 241 (1985), S. 67-76 
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Pars anterior ; Cell differentiation ; Grafts ; Mesenchymal cells ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Comparative studies have been made of development of the adenohypophysis using the Rathke's pouch (RP)-derived model system. Rathke's pouch with associated mesenchyme and ventral hypothalamus, was microsurgically isolated from 15-day fetal rats and placed in mild trypsin solution. Three variations of donor tissue were isolated and transplanted beneath the kidney capsule of adult hosts: A) pure pouch epithelium; B) pouch epithelium plus mesenchyme; and C) pouch epithelium with mesenchyme and ventral hypothalamus. After 30 days the grafts were isolated and processed for light and electron microscopy. Cell types were characterized by immunostaining as well as by morphological criteria. In group A well differentiated mammotrophs dominated the grafts, many of which were hypertrophied with widely dilated endoplasmic reticulum and Golgi saccules. Mammotrophs, frequently with mitotic figures, were distributed evenly throughout the grafts. Somatotrophs and gonadotrophs were neither abundant nor well differentiated in group A, but were both abundant and more extensively differentiated in groups B and C. Both somatotrophs and gonadotrophs were typically localized at margins of the graft adjacent to connective tissue spaces. Well differentiated mammotrophs were present in groups B and C although there were fewer hypertrophied mammotrophs than in group A; and immunoreaction to prolactin was weaker than in group A. Tumor-like features found in all three groups included some loss of tissue integrity and large, vascular lakes unlined by endothelium. These findings suggest that differentiation of mammotrophs may be inhibited in part by mesenchyme associated with Rathke's pouch, since in the absence thereof these cells become hyperplastic. Conversely, differentiation of somatotrophs and gonadotrophs appears more dependent on these mesenchymal elements for normal development.
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