Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.24.3 (collagenase)
18,340 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

One obstacle to the clinical implementation of endothelial cell seeding of vascular prostheses is the difficulty in derivation of large numbers of autologous endothelial cells from blood vessels of patients requiring vascular grafting. Capillary endothelial cells obtained from fat have been suggested as an abundant alternative to large-vessel endothelium for graft seeding. The object of this study was to evaluate the performance of 4-mm internal diameter (ID) Dacron Microvel grafts seeded with omentally derived microvascular endothelial cells. Six-cm lengths of the test grafts were implanted bilaterally into canine carotid arteries. One of each pair of grafts was seeded with endothelial cells (means = 8.4 x 10(6)) derived from collagenase digestion of autologous omental fat samples. The contralateral graft of each pair was nonseeded. At 5 weeks postoperatively, the grafts were harvested and evaluated. The mean patencies of both the seeded and nonseeded grafts were 89 percent. The mean thrombus-free surface area for seeded grafts was 95 +/- 11 percent. This value was significantly different statistically from the mean thrombus-free surface area of nonseeded grafts, which was 43 +/- 19 percent (P less than .05). Histologically, midgraft regions of seeded grafts were cellular, stained positive for collagen, and were characterized by inner capsules ranging in thickness between 35-94 microns. Luminal cells were identified as endothelial by peroxidase antiperoxidase staining techniques. Midgraft regions of nonseeded grafts demonstrated thrombus accumulation, limited cellularity, and inner capsules between 59-194 microns thick. Scanning electron microscopy of seeded grafts revealed smooth luminal surfaces with tight junctions between adjacent cells; surface cells were not present on midgraft regions of nonseeded grafts. In conclusion, endothelial cells derived from omental fat successfully surfaced on Dacron grafts and imparted characteristics to the graft that would predict long-term graft success.
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PMID:Microvascular endothelial cell seeding of small-diameter Dacron vascular grafts. 297 83

Cultured mammary cells depend on interaction with a substratum for functional differentiation, even in the presence of lactogenic hormones. Protein synthesis and secretion by mouse mammary epithelial cells on floating collagen gels and (EHS) matrix were compared. Cells were prepared by collagenase digestion of tissue from mid-pregnant mice. Protein synthesis was consistently greater in cells attached to EHS matrix, and was associated with proportionately higher rates of protein secretion into culture medium. Cells on EHS secreted protein into a luminal space formed within multicellular alveolus-like structures. Luminal secreted protein, extracted by EGTA treatment of cells in situ, constituted up to 40% of total secreted radiolabeled protein for cells on EHS matrix. The EGTA extract contained a higher proportion of casein and lactoferrin, whereas transferrin was predominantly in the medium. This indicated that cells on EHS matrix had become polarized and were secreting proteins vectorially. In contrast, EGTA treatment of cells on floating collagen gels released virtually no radiolabeled protein, showing that mammosphere formation was a property of cells on EHS. These biochemical observations were supported by ultrastructural evidence. In EHS cultures, the proportion of secreted protein in the luminal fraction, but not the distribution of secreted proteins, changed with time. This suggests that there may be leakage out of the lumen, or intraluminal degradation of protein after secretion. Nevertheless, the results suggest that cellular organization into mammospheres on EHS matrix promotes synthetic and secretory activity. This system provides a useful model for investigation of the regulation of milk secretion.
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PMID:Extracellular matrix and mouse mammary cell function: comparison of substrata in culture. 798 41

Visualization of water transport in cells, tissues and organs is an important, yet still difficult, task in morphological science. By using confocal microscopy and the fluid-phase fluorescent tracer technique, we visualized water secretion and estimated the routes of water transport across the acinar epithelia in rat parotid and submandibular glands. Confocal microscopy of whole glands perfused arterially with Lucifer yellow revealed a bright fluorescence at the basolateral space of acini. Luminal space was devoid of fluorescence, but revealed it after isoproterenol pretreatment, ductal infusion of fluorescent dextrans into the lumen, or tissue dissociation by collagenase. Under these conditions, stimulation of fluid secretion with carbachol caused a rapid decline of the luminal fluorescence intensity, indicating that the secreted water washed out the fluorescent probes in the acinar lumen. In the stimulated dissociated acini, the luminal fluorescence disappeared by 15 sec, but reappeared at 30-45 sec to maintain a low plateau level. By assuming that the tight junction was 'paralyzed' by the collagenase digestion and that the paracellular fluid transport could not influence the dilution of Lucifer yellow, we estimated that the initial water secretion by CCh occurs via the transcellular pathway, while later than 30-45 sec the additional water permeates through the paracellular pathway.
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PMID:Visualization of 'water secretion' by confocal microscopy in rat salivary glands: possible distinction of para- and transcellular pathway. 1456 2

Cultured mammary cells depend on interaction with a substratum for functional differentiation, even in the presence of lactogenic hormones. Protein synthesis and secretion by mouse mammary epithelial cells on floating collagen gels and (EHS) matrix were compared. Cells were prepared by collagenase digestion of tissue from mid-pregnant mice. Protein synthesis was consistently greater in cells attached to EHS matrix, and was associated with proportionately higher rates of protein secretion into culture medium. Cells on EHS secreted protein into a luminal space formed within multicellular alveoluslike structures. Luminal secreted protein, extracted by EGTA treatment of cells in situ, constituted up to 40% of total secreted radiolabeled protein for cells on EHS matrix. The EGTA extract contained a higher proportion of casein and lactoferrin, whereas transferrin was predominately in the medium. This indicated that cells on EHS matrix had become polarized and were secreting proteins vectorially. In contrast, EGTA treatment of cells on floating collagen gels released virtually no radiolabeled protein, showing that mammosphere formation was a property of cells on EHS. These biochemical observations were supported by ultrastructural evidence. In EHS cultures, the proportion of secreted protein in the luminal fraction, but not the distribution of secreted proteins, changed with time. This suggests that there may be leakage out of the lumen, or intraluminal degradation of protein after secretion. Nevertheless, the results suggest that cellular organization into mammospheres on EHS matrix promotes synthetic and secretory activity. This system provides a useful model for investigation of the regulation of milk secretion.
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PMID:Extracellular matrix and mouse mammary cell function: Comparison of substrata in culture. 2751 68