Of 1130 patients who had undergone a total of 1298 kidney transplantations, 40 developed a stenosis of the ureter (3.1%). In all of the transplants, the anastomosis between the ureter and the urinary bladder was established as extravesical ureteroneocystostomy. Up until 1984, in cases where an obstruction in the urinary tract was suspected, the diagnosis was made by an i.v. pyelogram or by nuclear scans of the transplant. Thereafter, in 28 patients, in cases of sonographically suspected stenosis of the ureter, the diagnosis was established by means of a simplified Whitaker test. Nearly one-half of the stenoses of the ureter developed within the first 3 months after transplantation. In five patients (12.5%) the stenosis developed significantly later, 3-10 years after kidney transplantation. Seventy percent of the stenoses were localized in the distal third of the ureter. About 75% of the surgically explored stenoses could be corrected by resection and reimplantation of the ureter. During the post-operative follow-up, restenosis occurred in three patients. At present, 62% of all patients whose ureteric stenoses were corrected have well-functioning kidney transplants.
In contrast to adherent cells, cells growing in suspension in up to 30% of transfected human primary T lymphocytes. and particularly hematopoietic cells, are notoriously difficultFlow cytometry analysis performed on T lymphocyte subto transfect in vitro using nonviral approaches. In the sets revealed that 8 and 9%, respectively, of CD4 and CD8 present study, the effect of cell adhesion on gene transfer cells could be transfected with a plasmid carrying the green efficacy was investigated by allowing hematopoietic cells fluorescent protein gene. Other adherent cells, such as to bind to an adherent cell monolayer (ACM) before being MS5 murine stromal cells or HeLa epithelial cells, were subjected to cationic liposome-mediated DNA transfer. also a compatible matrix for AAL. Moreover, the pCMV Human CD34 and T CD4 cell lines were cultivated on an plasmid was present in similar amounts in the nuclei of TF1 ACM constituted of murine fibroblast NIH3T3 cells and cells transfected in suspension or with the AAL procedure. transfected with a plasmid carrying the -galactosidase These data raise the possibility that cell matrix/ gene. X-gal staining showed that up to 27% of the cells hematopoietic cell interactions might govern expression of expressed the transgene. In contrast, less than 0.1% of the transgene in hematopoietic cells growing usually in these cells were positively transfected in suspension. This suspension, but not endocytosis of liposome/DNA particles adhesion-assisted lipofection (AAL) procedure was also and plasmid migration to the cell nucleus. successfully tested on blood lymphocytes, since it resulted
We investigated transfection rates of CD34+ haematopoietic progenitor cells (HPC) or haematopoietic cell lines (TF-1, KG1a and K562) using the LacZ gene as a reporter and cationic liposomes. The transfection efficiency of CD34+ haematopoietic progenitor cells (HPC) or TF-1, KG1a and K562 grown in suspension is very low (average percentage of 0.013 for HPC and 0.03 for cell lines). Adhesion of HPC or cell lines to plates by immunological or physical methods significantly enhances transfection efficiency; however, the percentage of transfected cells still remained low. We found that adhesion of TF-1, KG1a and K562 HC to MS-5 stroma cells or NIH-3T3 fibroblast cells increased transfection efficiency. Under these conditions transfection is achieved in 11.2-25% (mean 18.30%) for the cell lines and 13.6% (range 8.2-24.2%) for CD34+ HPC. These results indicate that liposome-mediated transfection of HC is significantly increased when cells are grown in adherence to stroma or fibroblast monolayers.
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