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

Transfection is a technique for inducing transformation of normal fibroblasts (NIH 3T3) with DNA (oncogenes) from human tumors. Our goal was to determine if these transformed cells expressed antigens associated with malignancy. NIH 3T3 cells were transfected with DNA fragments from a human acute lymphocytic leukemia (ALL 1-69), and transformed colonies were selected for growth in soft agar. Transfected cells containing human DNA sequences demonstrated by Southern blot analysis were used to immunize Balb/C mice. Monoclonal antibodies were produced and screened for binding to the parental leukemia (ALL 1-69), transfectant (17(2], and 3T3 cells in an enzyme-linked assay. A monoclonal antibody (IgM kappa) designated 17-9H3 bound to ALL 1-69 and secondary transfectant 17(2) but not to NIH 3T3 plasma membranes. Immunoperoxidase staining confirmed this binding pattern and demonstrated that the antigen was expressed on the cell surface. Expression of the antigen by transfectants directly correlated with the presence of a single 6.1 kilobase human DNA sequence. The antibody binding site of the antigen was inactivated by trypsin, glucosidase, and hyaluronidase. Binding of the 17-9H3 antibody was selective for acute lymphocytic leukemias (5/8) and osteogenic sarcomas (33/36), although other tumor types did demonstrate significant binding by immunoperoxidase staining. The majority of normal tissues did not bind 17-9H3 with the exception of some metabolically active cells (renal tubular epithelium, secretory epithelial cells, and cardiac smooth muscle), germ cells, Leydig cells of the testes, and some lymphoid cells. Monoclonal antibodies to oncogene-associated antigens may be potentially useful for cancer diagnosis and therapy and as probes for oncogene isolation.
 ...
PMID:A novel approach to production of antitumor monoclonal antibodies: antibody to a cell surface glycoprotein associated with transformation by a human oncogene. 637 59

Several techniques have been devised for the dissociation of tissues for primary culture. These techniques can affect the quantity and quality of the isolated cells. The aim of our study was to develop the most appropriate method for the isolation of human umbilical cord-derived mesenchymal (hUCM) cells. In the present study, we compared four methods for the isolation of hUCM cells: three enzymatic methods; collagenase/hyaluronidase/trypsin (CHT), collagenase/trypsin (CT) and trypsin (Trp), and an explant culture (Exp) method. The trypan blue dye exclusion test, the water-soluble tetrazolium salt-1 (WST-1) assay, flow cytometry, alkaline phosphatase activity and histochemical staining were used to evaluate the results of the different methods. The hUCM cells were successfully isolated by all methods but the isolation method used profoundly altered the cell number and proliferation capacity of the isolated cells. The cells were successfully differentiated into adipogenic and osteogenic lineages and alkaline phosphatase activity was detected in the hUCM cell colonies of all groups. Flow cytometry analysis revealed that CD44, CD73, CD90 and CD105 were expressed in all groups, while CD34 and CD45 were not expressed. The expression of C-kit in the enzymatic groups was higher than in the explant group, while the expression of Oct-4 was higher in the CT group compared to the other groups. We concluded that the collagenase/trypsin method of cell isolation yields a higher cell density than the others. These cells expressed a higher rate of pluripotent cell markers such as C-kit and Oct-4, while the explant method of cell isolation resulted in a higher cell proliferation rate and activity compared to the other methods.
 ...
PMID:Comparison of different methods for the isolation of mesenchymal stem cells from human umbilical cord Wharton's jelly. 2227 9

Scaffold-based delivery of bioactive molecules capable of directing stem cell differentiation is critical to the development of point-of-care cell therapy for orthopedic repair. Dexamethasone-conjugated hyaluronic acid (HA-DXM) was synthesized and combined with hydrolytically degradable, photo-cross-linkable PEG-bis(2-acryloyloxy propanoate) (PEG-bis-AP) to form semi-IPNs. Dexamethasone (DX) release was limited in physiological buffer and substantially increased in the presence of encapsulated human mesenchymal stem cells (hMSCs) or exogenous hyaluronidase, confirming that release occurred primarily by a cell-mediated enzymatic mechanism. hMSCs encapsulated in PEG-bis-AP/HA-DXM semi-IPNs increased osteoblast-specific gene expression, alkaline phosphatase activity, and matrix mineralization, attaining levels that were not significantly different from positive controls consisting of hMSCs in PEG-bis-AP/native HA cultured with DX supplementation in the culture medium. These studies demonstrate that PEG-bis-AP/HA-DXM semi-IPNs can provide cell-mediated release of bioactive free DX that induces hMSC osteogenic differentiation. This approach offers an efficient system for local delivery of osteogenic molecules empowering point of care applications.
...
PMID:Cell-Mediated Dexamethasone Release from Semi-IPNs Stimulates Osteogenic Differentiation of Encapsulated Mesenchymal Stem Cells. 2625 27