References

E., Researches on the ultracellular digestion of invertebrates, J. Microscop. Sci, 24, 155, 1884. 2. Cohn, Z. A., The fate of bacteria within the phagocytic cells. II. The modification of intracellular degradation, J. Exp. Med., 117, 43, 1963. 3. Fornusek L. and Vetvicka, V., Immune System Accessory Cells, CRC Press, Boca Raton, 1992. 4. van Furth, R., Identification of mononuclear phagocytes overview and definitions, in Reagents for Studying Mononuclear Phagocytes, Adams, D....

B lymphocyte cloning

Proliferation of pre-B cell colonies in III. Clonal assay for murine lymphohematopoietic progenitors 81 IV. Clonal assay for fetal V. Long-term bone marrow A. Dexter myeloid B. Whitlock-Witte lymphoid C. Establishment of human bone marrow stromal cells 86 D. Growth of human B cell E. Isolation of B lineage precursor F. Mouse stromal Growth and differentiation of hematopoietic cells are extremely complex processes involving interaction of numerous cell types...

Phagocytosis

The term phagocytosis comes from the Greek phagein, meaning to eat, and is used to describe the uptake of solid particular material such as bacteria, erythrocytes, viruses, fungi, and other organic and inorganic materials. Although the ingestion of foreign material by cells was observed and mentioned by several earlier scientists e.g., Koch and Roser described cells filled with anthrax bacilli, but the significance eluded them31 it was first described by Metchnikoff more than 100 years ago.1...

Pinocytosis

Pinocytosis was originally discovered by Haeckel43 in Tethys fibria, but not until 1930 was the microscopy of single cells advanced sufficiently to allow Lewis44 to describe the uptake of extracellular fluid in full detail. The name pinocytosis, from the Greek pinos, meaning to drink, describes the uptake of anything from droplets of fluid, coloids, or immunocomplexes to soluble macromolecules. As a rule, pinocytic vesicle is always filled with fluid. Two major types of pinocytosis have been...

Protocol

Note All materials and reagents must be sterile and proper aseptic technique must be used when handling the cells. 1. Prepare MEM medium with 2.1 methylcellulose solution. Aliquot into 100-ml lots and store at -30 C. 2. Prepare MEM medium with 15 FCS, 5 x 10-5 M 2-mercaptoethanol, IL-7, and 0.8 methylcellulose use 40 ml of 2.1 stock medium per 100 ml of final medium . 3. Isolate bone marrow cells see Chapter 1 and prepare cell suspension. 4. Mix 3 x 105 of cells plate in 0.1 ml with 2.9 ml of...

Mishell Dutton balanced salt solution

Prepare both stock solutions and sterilize them by filtration through 0.22 m filter. Store at 4 C for no longer than 2 months. 2. Before experiment, prepare working solution Add 100 ml stock solution 1 to 700 ml H2O and then add 100 ml stock solution 2. Add H2O to 1000 ml and use NaOH to keep pH at 7.4.

Enrichment of human basophils

Basophils are the least abundant of the circulating leukocytes, and thus are relatively difficult to purify. Isolation protocols based on density-gradient centrifugation have been reported to result in basophil enrichments of up to 50 , although significant contamination with other cell types, particularly lymphocytes, still exists.1 One key consideration during the enrichment of basophils is preventing their stimulation and degranulation. The method described is based on the report of Leonard...

Isolation and characterization of neutrophils

Percoll sedimentation and Ficoll-Paque II. Magnetic cell sorting with anti-CD15 III. Functional A. Measurement of phagocytic B. Microbicidal C. Nitroblue tetrazolium dye reduction D. Superoxide anion O2- E. Hydrogen peroxide Neutrophils or polymorphonuclear leukocytes PMNs are the most abundant type of leukocyte in normal peripheral blood. They participate in the effector phase of immune responses, playing important roles in inflammation and in the pathogenesis of a number of diseases.1,2...

The Authors

Rafael Fernandez-Botran, Ph.D., is an associate professor in the Department of Pathology and Laboratory Medicine, Division of Experimental Immunology and Immunopathology, School of Medicine, University of Louisville, Kentucky. Dr. Fernandez-Botran received his bachelor of science degree in biological chemistry from San Carlos University, Guatemala in 1979 and his Ph.D. in microbiology from the University of Kansas Medical Center, Kansas City in 1985. Dr. Fernandez-Botran worked as a...