We describe a system for analysis of the intracellular pathways in the biosynthesis and packaging of functionally important proteins in human myeloid cells. The human promyelocytic cell line HL-60 was used since peripheral blood neutrophils are terminally differentiated and do not actively synthesize protein. Cells were disrupted by nitrogen cavitation and subcellular organelles in postnuclear ...

The present study was designed to compare fertilization rates following oocyte exposure to spermatozoa from different buffalo bulls, using three sperm preparation methods i.e. percoll density gradient, swim-up (modified Ca free Tyrode’s medium and TALP medium) and sodium citrate washing prior to co-incubation with oocytes. Buffalo oocytes were aspirated from ovarian follicles within 1.5 to 2 ho...

Previous studies on human eosinophils often have used cells from patients with hypereosinophilia syndrome or parasitosis owing to the difficulty in isolating pure populations of eosinophils from normal individuals. In the present study, human eosinophils were isolated with a purity of 97%, with 70% recovery from normal individuals with blood eosinophil counts of less than 3%. Human eosinophils ...

The isolation of pancreatic islets from collagenase-digested Wistar rat pancreas was shown by the sedimentation method at a unit gravity using Percoll solution with a density of 1.041 g/ml. The density of digested exocrine tissues was in the range of 1.013-1.041 g/ml, while that of purely isolated islets was in the narrow range of 1.066-1.075 g/ml. More than a hundred islets were obtained freel...

BACKGROUND The purpose of this study was to optimize the electrophoretic conditions that should be used for the effective isolation of functional human spermatozoa and to determine whether this method of isolating cells was associated with oxidative stress and DNA damage. METHODS Human spermatozoa were prepared by repeated centrifugation, discontinuous density gradient centrifugation and elec...

BACKGROUND Because few definitive markers are available for hepatic cancer stem cells (HCSCs), based on physical rather than immunochemical properties, we applied a novel method to enrich HCSCs. METHODOLOGY After hepatic tumor cells (HTCs) were first isolated from diethylinitrosamine-induced F344 rat HCC model using percoll discontinuous gradient centrifugation (PDGC) and purified via differe...

Peripheral blood stem cells (PBSC) were collected for autotransplantation by a total of 46 continuous-flow leukaphereses in 17 children with various types of cancer in whom the stem-cell pool had been expanded by chemotherapy. As the cells collected by leukapheresis were contaminated with many visible cell clumps, platelets, and erythrocytes, they were separated from the platelet-rich plasma (P...

The prespore vesicle (PSV) is an organelle which secretes spore coat proteins and gal/galNAc polysaccharides from prespore cells of Dictyostelium. By combining the techniques of protein A-gold immunocytochemistry and ricin-gold affinity cytochemistry we have demonstrated colocalization of the lysosomal enzyme alpha-mannosidase with gal/galNAc polysaccharides in prespore vesicles and the spore c...