Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: DrugBank:EXPT01586 (G418)
 2,237 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The skeletal muscle capillary bed may be an ideal recipient site for transplantation of genetically modified autologous endothelial cells and thus provide a basis for a technique of somatic gene therapy that would be applicable to a variety of acquired and inherited human diseases. The purpose of this study was to test the hypothesis that adhesion of lac-Z-transduced microvascular endothelial cells (MVEC) in the skeletal muscle capillary bed in vivo is dependent on the duration of arterial occlusion after injection of the transduced MVEC. MVEC derived from the abdominal fat pad of syngeneic rats (Wistar F-455) were transfected with the BAG vector, a replication-incompetent retroviral vector containing the lac-Z gene for beta-galactosidase and the Tn5 gene for selection of the transduced cells by the neomycin analogue, G418. lac-Z-transduced MVEC were radiolabeled with 125I-PKH-95, and, after the femoral artery was occluded for 10 min, these cells (1 to 2 x 10(6)) were injected intraarterially into the rat hindlimb. In the experimental groups the femoral artery clamp was removed at 0, 60, or 120 min after injection. A control group without pre- or postinjection femoral arterial occlusion was also studied. Adhesion of MVEC in the skeletal muscle capillary bed (mean percentage of injected 125I activity) was determined in groups of 4 rats at 1 day, 1 week, and 1 month after injection. Adhesion of the transduced MVEC did not increase as the duration of femoral artery occlusion after injection was increased.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Transplantation of lac-Z-transduced microvascular endothelial cells into the skeletal muscle capillary bed of the rat hindlimb occurs independent of the duration of femoral artery occlusion after injection of cells. 799 42

Cellular proliferation is regulated in both positive and negative ways. However, direct selection for growth inhibitory control elements is limited by the difficulty in identifying a growth inhibited cell against a background of cells which are proliferating. This study describes a positive selection technique for growth inhibited cells. This method is based on the retention of a lipophilic fluorescent dye which nonspecifically labels plasma membranes and distributes between daughter cells with membrane lipid as cells proliferate. Characterization of this assay is described using an epithelial cell line which is growth inhibited in response to transforming growth factor beta (TGF-beta) and dexamethasone and several mutant clones of that line which lack responsiveness to TGF-beta. Retention of dye in response to the growth inhibitors is proportional to the inhibition of thymidine incorporation of those cells. Mixing experiments were also carried out in which G418 resistant TGF-beta responsive epithelial cells were mixed with TGF-beta nonresponsive mutants. The mixture was labeled with PKH-2 and exposed to TGF-beta for 3 days. Subsequently, consecutive fractions of cells sorted on the basis of fluorescence intensity were selected in G418, and the TGF-beta responsive epithelial cells were found predominantly in the most fluorescent cells in the population. This method provides a positive selection for growth inhibited cells which may, in combination with classical gene transfer techniques, provide a way to select for growth inhibitory genes in a manner analogous to the focus forming assay selection for oncogenes.
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PMID:Identification of growth inhibited cells by retention of a lipophilic fluorescent dye. 824 Oct 26