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Epithelial-cell enriched primary cultures have been established from rat ventral prostate (RVP). Minced ventral prostates were dissociated with 0.5% collagenase in F12K tissue culture medium containing 1% fetal bovine serum. This treatment resulted in the gradual removal of stromal elements from the base of the epithelial cells. After 60 minutes of digestion the aggregates of epithelial cells were washed and plated at high density in F12K plus 10% horse serum. After 48 hours in vitro the unattached cells were removed from the culture dishes, washed, and reinoculated into new culture vessels containing fresh medium. After 96 hours in vitro, the aggregates had attached to the culture vessels and spread out to yield discrete patches of epithelial cells. By 144 hours in vitro the patches of cells had grown and coalesced to form a semi-confluent monolayer of epithelial cells. Ultrastructrual examination of these cultures indicated that adjacent cells were joined by desmosomes and tight junctions and had formed "lumen-like structures" into which projected microvilli. In addition, the cells contained secretory granules and tonofilaments, giving them a morphological appearance similar to prostate epithelial cells in the intact organ. The primary cultures also retained histochemical activities for acid phosphatase, beta-glucuronidase, and succinic dehydrogenase that were similar to the intact organ.
Anat Rec 1980 Jun
PMID:Isolation, culture and characterization of epithelial cells derived from rat ventral prostate. 625 35

The secretory acinar cells of parotid glands from rats of varying ages have been examined by electron microscopy to determine what age-related changes occur in these cells. The most prominent change noted in these cells is the progressive increase in the amount of lipofuscin granules with age. Lipofuscin granules are membrane-bound structures consisting of lipids, other subcomponents, and a matrix. In addition, these cells contain lipid droplets that are not associated with any other components and tend to accumulate at the base of the cells in older rats. Also, many acinar cells in the glands of old rats contain altered secretory granules which appear to be in the process of degeneration. The accumulation of lipid and degenerating secretory granules appears to be related to the reduced level of cellular secretory activity in the glands of older rats. It is possible that these two types of inclusions contribute to the formation of lipofuscin granules. Lipofuscin and degenerating secretory granules are associated with acid phosphatase, which is demonstrated cytochemically, indicating that these granules are lysosomal structures.
Anat Rec 1984 Jul
PMID:Changes in the secretory acinar cells of the rat parotid gland during aging. 646 43

Fetal rabbits (days 13-32), rats (days 14-22), and hamsters (days 11-15) and selected postnatal animals were examined for pulmonary macrophages or their precursors in 2-micron sections stained by PAS-lead hematoxylin (all species), electron micrographs (rabbit and rat), and cytochemical incubations for acid phosphatase (rabbit and rat), aliesterase, and N-acetyl glucosaminidase (rabbits). All methods revealed macrophages in perinatal specimens. The appearance and distribution of these cells were compared in the different preparations to establish the reliability of PAS-lead hematoxylin for identifying them in less developed fetal lungs, where they are less active for lysosomal enzymes the earlier the stage examined. In the sections, macrophages are seen to possess a round or indented nucleus, an irregular contour, and a deep purplish-gray cytoplasm containing a variety of pink PAS-stained granules, equated with heterolysosomes by ultrastructural cytochemistry. In less developed lungs, macrophages occur along with putative precursors having a more rounded outline and fewer PAS-stained granules. In pseudoglandular lungs these precursors predominate over rather vacuolated macrophages resembling Hofbauer cells. In all three species both cell types first appear in the stroma during the bronchial bud stage and are frequently seen to divide from that time on. The earliest precursors have a relatively sparse cytoplasm which later increases in daughter cells. Hofbauer-like cells disappear during the canalicular stage of development, replaced by macrophages and transitional forms from the more rounded precursors. In day 21 rabbit lungs, scattered stromal cells are reactive for aliesterase, and, some days later, for acid phosphatase and glucosaminidase. Free mononuclear cells are rare in airways of pseudoglandular lungs but become common later. A day or two before birth in rats, free cells range between rather undifferentiated leukocytes to typical macrophages, but cells with the macrophage's complete repertory of inclusions are seen only after birth. In the fetus, typical monocytes were not identified in either the pulmonary stroma or the airways. A replicating population of macrophage-like cells therefore resides in fetal lungs. It is established before bone marrow is formed and, in rats, before monocytes have appeared in the circulation.
Anat Rec 1984 Jan
PMID:Development of macrophages in the lungs of fetal rabbits, rats, and hamsters. 671 32

The cell surface characteristics of degenerating cells and phagocytes, as well as the participation of lysosomes in the cell death process associated with the early embryogenesis of chick lens rudiment, were studied by means of scanning electron microscopy and cytochemically using the Gomori-beta-glycerophosphate method for acid phosphatase. The prospective dying columnar epithelial cells lose their apical and basal processes and become rounded. The rounded, isolated, dying cells initially show a rough surface with some cytoplasmic constrictions followed by progressive break-up into several pitted fragments. Coincident with the loss of the columnar cell shape, acid phosphatase is localized within the Golgi apparatus and autophagic vacuoles which progressively increase in size. In contrast, the isolated dying cells and fragments do not show significant acid phosphatase activity. The role of lysosomes in this degenerative process is discussed. Neighboring epithelial cells phagocytose the dead cell fragments, becoming nonspecialized phagocytes. These consist of columnar epithelial cells and free cells which have migrated from the lens epithelium. Two mechanisms of internalization are observed. The most frequent mechanism takes place in both the columnar epithelial cells and the free cells, and consists of the progressive engulfment of the fragments into craters of the cell surface. The other mechanism is only detected in the free cells and takes place by pseudopod engulfment. We suggest that both phagocytic procedures could be related to the degree of intercellular connection. The presence of phagocytic internalization by crater formation in the epithelial cells could be a mechanism preserving the epithelial stability, which is necessary for a normal morphogenesis. Small microprocesses binding the surface of the phagocyte and the fragment are present prior to the internalization process. In the lens stalk and in the space located between the ectoderm and the lens vesicle, there are some cells displaying migratory characteristics. This fact suggests that an active migration of epithelial cells from the lens stalk could account for the process of detachment of lens vesicle from the ectoderm. The free cells appear to undergo an in situ progressive degeneration.
Anat Rec 1984 Jan
PMID:The mechanisms of cell death and phagocytosis in the early chick lens morphogenesis: a scanning electron microscopy and cytochemical approach. 671 33

Guinea pig polymorphonuclear leukocytes (PMNs), rich in glycogen granules, were collected from sodium-caseinate-induced peritoneal exudate. When these cells were incubated with rickettsiae, many microorganisms were phagocytized within 30 minutes at 35 degrees C and vacuoles up to 5 microns in diameter containing glycogen granules were present. Contained within these vacuoles were phagocytized extracellular material and a dense, lysosomelike substance that was acid phosphatase positive. These vacuoles, which were interpreted to be autophagosomes, were absent from PMNs that had not been stimulated with microorganisms. The number of rickettsiae in the PMN did not appear to be related to the number of autophagosomes. About 8% and 80% of thin-sectioned profiles of PMNs contained these vacuoles after 30 minutes and 4 hours incubation, respectively. After 4 hours, the PMNs contained multiple autophagosomes. Almost all of the glycogen granules were in autophagosomes in some of the cells. In some PMNs, discontinuous membranes encircled some glycogen. When PMNs were initially incubated with thorium dioxide and ferritin, and extensively washed prior to incubation with rickettsiae, glycogen was found surrounded by flattened secondary lysosomes containing the dense tracers. Some autophagosomes also contained the electron-dense tracers. These results suggest that rickettsiae induce the rapid formation of glycogen-containing autophagosomes in guinea pig peritoneal PMNs in vitro.
Anat Rec 1984 Mar
PMID:Glycogen autophagosomes in polymorphonuclear leukocytes induced by rickettsiae. 672 Dec 27

This study has examined the fine structure and some cytochemical characteristics of the endodermal and mesothelial cells of the rhesus monkey yolk sac between 25 and 66 days of gestation. The endodermal cells were characterized by abundant granular endoplasmic reticulum, some agranular endoplasmic reticulum, a well-developed Golgi apparatus, and numerous large mitochondria. During the earlier part of the period studied, endodermal cells had a few acid phosphatase and arylsulfatase-positive lysosomes and moderate numbers of catalase-positive microperoxisomes. During the later stages of development, large granules (believed to be lysosomes) with a heterogeneous content were numerous in the cytoplasm. Mesothelial cells showed fewer development changes. Throughout this period they were usually flattened cells with long microvilli, small mitochondria, and limited amounts of granular endoplasmic reticulum. The mesothelial cells had acid phosphatase reaction product in the Golgi region and occasional large vesicles, but were negative for arylsulfatase and catalase. One specimen was incubated at 37 degrees C in the presence of horseradish peroxidase in order to examine endocytosis. Both the mesothelial cells and endodermal cells internalized the peroxidase into a variety of cytoplasmic vesicles. Based on their cytology, the endodermal cells may function in the synthesis of serum proteins during this period, as has been suggested in other species. They may also be involved in lipid metabolism. The mesothelial cells appeared less synthetically active, but evidence suggested that they may be involved in collagen and extracellular matrix production. The endocytic activity displayed by both cell types may indicate a role in fluid and metabolite transfer across the epithelia. The cytology of both cell types was very similar to that described for human yolk sacs, suggesting that the rhesus monkey may be a useful species in which to study the maturation of yolk sac function.
Anat Rec 1983 Feb
PMID:A fine structural and cytochemical study of the rhesus monkey yolk sac: endoderm and mesothelium. 684 66

Little information is available concerning enzyme activity in primordial germ cells (PGCs) of the early chick embryo. The present study is designed to examine the disposition of alkaline and acid phosphatase activity in the PGCs during their migration into the developing gonads of the early chick embryo. White Leghorn chick embryos were sacrificed at daily intervals from 1 to 6 days of incubation. Following sacrifice the embryos were fixed, dehydrated, and embedded in glycol methacrylate (GMA). Alkaline and acid phosphatases were demonstrated by the simultaneous diazo-coupling method. The embryonic tissues at the different ages were examined for PGCs and the histochemical reactions for alkaline and acid phosphatases in these cells evaluated. Acid phosphatase activity did not appear within PGCs until 3 days of incubation, and then in only a few PGCs in the active phase of their migration in the dorsal mesentery, suggesting that there is no large wave of degeneration of these cells during migration. Alkaline phosphatase activity was observed as early as 2 days of incubation in PGCs during the passive phase of their migration in extraembryonic blood vessels. Alkaline phosphatase-positive PGCs in the active phase of migration were also found in the dorsal mesentery; however, the cellular localization of this enzyme differed from that observed in the passively migrating PCGs, indicating that there are alterations in the metabolic activities of these cells during the active and passive phases of migration.
Anat Rec 1982 Mar
PMID:Acid and alkaline phosphatase activity in migrating primordial germ cells of the early chick embryo. 707 83

Resorption of uncalcified cartilage in the embryonic chick femur appears to be mediated by two types of mononuclear cells. One cell type lies flattened and adherent along the surface of the cartilage matrix into which it extends cellular processes. Cytological characteristics of a large, euchromatic nucleus containing a nucleolus, and cytoplasm containing moderate to extensive amounts of rough endoplasmic reticulum indicate that these are protein synthetic cells. Macrophages, characterized by a pleomorphic shape and cytoplasm containing numerous mitochondria and vesicles, comprise the second cell type. These may be seen lying in contact with cartilage matrix, but are more likely located in the nonhematopoietic marrow adjacent to resorbing cartilage, where they establish close cellular associations with protein synthetic cells. Alkaline and acid phosphatase histochemical studies differentiate these two cellular types. Marrow alkaline phosphatase activity is restricted to the cartilage-marrow interface from which it diffuses a short distance into cartilage matrix, but does not diffuse into nearby marrow. Intracellular alkaline phosphatase is present only in protein synthetic cells that line the surface of cartilage, and thus appears to be produced by these cells. Acid phosphatase positive macrophages are scattered throughout the marrow, but are found in greatest concentrations in the region of cartilage resorption. They are rarely in direct contact with cartilage, and there is no evidence that acid phosphatase is released from these cells. The relative localizations and the presence of cellular interactions of these two cell types suggests that protein synthetic cells may be of fibroblastic origin, and may play a primary role in cartilage degradation, while macrophages, in keeping with biochemical evidence, play an adjunct or possibly a regulative role.
Anat Rec 1982 Apr
PMID:The cellular organization of fibroblastic cells and macrophages at regions of uncalcified cartilage resorption in the embryonic chick femur as revealed by alkaline and acid phosphatase histochemistry. 707 91

Ultrastructural and cytochemical methods were utilized to study the human Fallopian tube fimbrial epithelium during the different stages of the menstrual cycle. Alkaline phosphatase reaction product was located along the apical and lateral plasma membranes of the secretory cells only, regardless of the stage of the cycle. The ciliated cells were almost devoid of any reaction product at all stages of the cycle. Acid phosphatase reaction product depicted the lysosomes. These appeared as electron-dense bodies, of almost equal numbers in the ciliated and the secretory cells at all stages of the cycle. Thus the number of lysosomes did not vary appreciable during the different stages of the menstrual cycle. Many lipid droplets were found in both cells; these were rimmed by acid phosphatase reaction product, and some were partially enveloped by electron-dense bodies containing acid phosphatase deposits. Acid phosphatase deposits were also found on the inner face of Golgi vesicles.
Anat Rec 1982 May
PMID:Ultrastructural localization of alkaline and acid phosphatases in the human fallopian tube epithelium during the menstrual cycle. 710 26

Osteoclasts residing on rims and walls of bony concavities on remodeling proximal tibia from growing rats were examined by light microscopy following stripping of the periosteal connective tissue layer. Comparison of these cells in situ and after transfer to glass slides revealed the presence of numerous mononucleate osteoclasts, as well as typical multinucleate forms, all exhibiting a ruffled border and acid phosphatase and succinic dehydrogenase activity. Macrophage-like cells were situated adjacent to osteoclasts in situ. Osteoblasts were relatively inconspicuous. The possibility that the basic functional osteoclast unit is a mononucleate cell is discussed.
Anat Rec 1982 Jun
PMID:A comparative study of osteoclasts: in situ versus smear specimens. 711 95


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