Lymphocytes, Animals, Mice, Inbred C57BL, Mice, Inbred DBA, Sheep, Humans, Mice, Silicon Dioxide, Immunoglobulin M, Colloids, Centrifugation, Density Gradient, Cell Separation, Cytotoxicity, Immunologic, Osmolar Concentration, Male, Immunology, Opthalmology and Optometry
Abstract
This report describes a unique modification of an isopycnic density gradient system utilizing as a separating menstrum colloidal silica (Ludox AM). The primary advantages of this preparation are: (1) It is chemically defined, allowing extremely reproducible cell separation employing different lots of material; (2) the physical parameters (pH, density, salt concentration) of the final gradient suspension can be manipulated over a wide range of values, allowing for the separation of many different biological materials; (3) it allows separation of very large numbers of lymphoid cells with greater than 95% recovery of applied cells; (4) separated cellular subpopulations can be easily washed free of silica and cellular function is retained. This paper is a report of the preparation and functional characteristics of the gradient material as it relates to the separation of very large numbers of lymphoid cell subpopulations in both mouse and man. Subpopulations of murine and human lymphocytes separated by this gradient material were assayed for IgM synthesis, T-cell mediated cytotoxicity, and lymphokine production.
Biochemistry and Cell Biology, Biomedical and Clinical Sciences, Biological Sciences, Animals, Cell Separation, Centrifugation, Density Gradient, Colloids, Cytotoxicity, Immunologic, Humans, Immunoglobulin M, Lymphocytes, Male, Mice, Mice, Inbred C57BL, Mice, Inbred DBA, Osmolar Concentration, Sheep, Silicon Dioxide, Immunology, Medical Microbiology, Opthalmology and Optometry, Biochemistry and cell biology
Abstract
This report describes a unique modification of an isopycnic density gradient system utilizing as a separating menstrum colloidal silica (Ludox AM). The primary advantages of this preparation are: (1) It is chemically defined, allowing extremely reproducible cell separation employing different lots of material; (2) the physical parameters (pH, density, salt concentration) of the final gradient suspension can be manipulated over a wide range of values, allowing for the separation of many different biological materials; (3) it allows separation of very large numbers of lymphoid cells with greater than 95% recovery of applied cells; (4) separated cellular subpopulations can be easily washed free of silica and cellular function is retained. This paper is a report of the preparation and functional characteristics of the gradient material as it relates to the separation of very large numbers of lymphoid cell subpopulations in both mouse and man. Subpopulations of murine and human lymphocytes separated by this gradient material were assayed for IgM synthesis, T-cell mediated cytotoxicity, and lymphokine production.
Published
1979
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