Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.2.1.36 (hyaluronidase)
 4,606 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A method is described for the isolation of biliary epithelial cells from rat livers by sequential treatment with EGTA, collagenase-hyaluronidase, trypsin, and deoxyribonuclease I and final separation on a discontinuous Percoll gradient using prekeratin antibodies as an immunohistochemical marker for these cells.
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PMID:The isolation of intrahepatic biliary epithelial cells from normal rat livers. 258 10

We describe a method for establishing the culture of bovine tracheal submucosal gland (BTG) cells, in which we have also examined the influence of a reconstituted basement membrane matrix derived from the Engelbreth-Holm-Swarm tumor (EHS) on the growth and morphological differentiation of these cells. BTG cells have been isolated by tissue enzymatic digestion using trypsin, deoxyribonuclease I, elastase, hyaluronidase and EGTA for 1 hr at 37 degrees C. Afterwards, cells and tissue were collected by centrifugation and were incubated for 15 min with 15% newborn calf serum to inactivate the proteolytic enzymes. Enzymatic digestion using only trypsin, centrifugation and inactivation steps were repeated three times. Using this protocol, we obtained 15 +/- 4 (X 10(6] cells per g of tracheal submucosa with 72 +/- 2% (n = 5) cell viability. On microscopic observation, isolated cells were mainly composed of serous type glandular cells. Cells were cultured in a 1:1 medium of Dulbecco's Modified Eagle's/Ham's F12 supplemented with 10% fetal calf serum and subcultured in either plastic flasks or flasks coated with EHS matrix. On the plastic, the BTG cells exhibited at confluency an epithelioid appearance. They stained positively with the immunofluorescent anticytokeratin antibody and contained PAS-staining granules. By electron microscopy, lactoferrin, a protein marker specific to the serous cells, was demonstrated immunocytochemically in small secretory vesicles. BTG cells cultured on EHS matrix revealed a significantly increased growth in comparison to those cultured on plastic. In post-confluent culture of BTG cells on EHS matrix, we observed numerous dome-like structures formed by differentiated cells which were joined together around luminal spaces.
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PMID:Growth and characterization of isolated bovine tracheal gland cells in culture. Influence of a reconstituted basement membrane matrix. 260 69

An inhibitory component that diminishes estrogen receptor (ER) binding to nuclei in vitro is present in cytosol prepared from calf uterus. The inhibitor is heat stable and resistant to enzymatic treatment with trypsin, chymotrypsin, proteinase K, deoxyribonuclease I, or ribonucleases A, T1, and U2. Results of chromatography on DEAE-cellulose and Sephadex G-150 indicate that the factor is a negatively charged macromolecule. Inhibitory activity is sensitive to sequential digestion with chondroitinase ABC, hyaluronidase, and heparinase. Approximately 70% of the inhibitory activity is destroyed by treatment with heparinase alone. Heparitinase destroys only 30% of this activity. Furthermore, the addition of pure hyaluronic acid or chondroitin sulfate to the ER-nuclei binding assay results in little inhibition, whereas addition of heparin inhibits 75% of receptor binding. Overall, these results indicate that glycosaminoglycans, present in bovine uterine cytosol, are capable of inhibiting ER-nuclei interactions. The most potent inhibitory glycosaminoglycan displays heparin-like characteristics.
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PMID:Characterization of a cytosolic inhibitor of calf estrogen receptor binding to nuclei. 330 79

Hyaluronic acid (HA), an abundant non-sulfated glycosaminoglycan component of the extracellular matrix, has applications in drug delivery, tissue engineering and as an ingredient in cosmetics. HA preparations containing high-molecular-weight polymers are also used in the treatment of inflammatory disorders such as arthritis and interstitial cystitis. Low-molecular-weight fragments derived from HA have been reported to induce pro-inflammatory cytokines such as IL-12 and TNF-alpha, and could therefore potentially exacerbate existing inflammation. We therefore examined the pro-inflammatory activity of HA preparations, since inflammatory reactions are known to occur following administration of HA. We tested low-molecular-weight fragments obtained from seven different HA preparations, either by sonication (approximately equals 3 x 10(5) Da) or by hyaluronidase digestion (approximately equals 1 x 10(4) Da), for the ability to induce the synthesis of IL-12 and TNF-alpha by human monocytic cells. We found that two of the seven HA preparations tested stimulated the synthesis of IL-12 and TNF-alpha by human monocytic cells. We unexpectedly found that the induction of IL-12 and TNF-alpha by these HA preparations was not due to their degradation to low-molecular-weight fragments, since their native high-molecular-weight forms possessed the same ability to stimulate IL-12 and TNF-alpha synthesis, but was due to the presence of contaminating DNA. Treatment of these two HA preparations with deoxyribonuclease I abrogated or reduced the induction of IL-12 and TNF-alpha. It is clear from this study that HA preparations can induce the synthesis of pro-inflammatory cytokines by monocytes. The ability of HA to act as a pro-inflammatory mediator may not, however, be related to the presence of low-molecular-weight HA fragments, but to the presence of DNA. The presence of pro-inflammatory DNA in HA preparations should be evaluated before its use, not only for the treatment of patients with inflammatory disorders, but also before many other applications.
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PMID:Pro-inflammatory activity of contaminating DNA in hyaluronic acid preparations. 1134 74

Normal human mammary tissue is composed of a glandular epithelium embedded within a fibrous and fatty stroma. Collagenase and hyaluronidase digestion of normal reduction mammoplasty specimens followed by differential centrifugation yields a suspension of single cells and cell aggregates that contain elements of the terminal ductal lobular units and stromal components of the mammary gland. The terminal ductal lobular units (TDLU) can be further dissociated to complete viable single-cell suspensions by treatment with trypsin, dispase II, and deoxyribonuclease I. These suspensions are suitable for cell separation and analysis in culture. Such studies indicate the existence of biologically distinct subpopulations of luminal-restricted, myoepithelial-restricted, and bipotent mammary epithelial progenitors detected by their ability to generate colonies of the corresponding progeny types in serum-free cultures. This review summarizes the methodology of the techniques required to generate and characterize the colonies obtained in vitro from these progenitors, as well as the special considerations and potential pitfalls associated with performing these protocols.
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PMID:Enzymatic dissociation and culture of normal human mammary tissue to detect progenitor activity. 1536 67