BIOMAT Database Logo BIOMAT Database Logo

Isolation of monocytes from whole blood by density gradient centrifugation and counter-current elutriation followed by cryopreservation: six years' experience. 2000

P K Lund, and G B Joø, and A B Westvik, and R Ovstebø, and P Kierulf
Department of Clinical Chemistry, The Research and Development Group, Ullevaal University Hospital, Oslo, Norway. perlund@online.no

BACKGROUND Monocyte purification by means of counter-current elutriation and subsequent cryopreservation for future use was initiated in 1986 and has been established as a routine since 1993. OBJECTIVE To sum up and evaluate our method for the isolation and preservation of monocytes. METHODS Peripheral blood mononuclear cells (PBMC) were isolated from healthy donor blood by density gradient centrifugation, and monocytes were isolated from the PBMC by counter-current elutriation centrifugation using the Beckman J-6M/E centrifuge. The monocytes were then cryopreserved at 135 degrees C and thawed when required for experimental use. RESULTS Results are given for the last 6 years, including 59 elutriations and the fractions containing monocytes. The mean purity of monocytes was 93% (range 64-98%); mean recovery was 51% (range 22-55%). Studies of CD14 expression and Annexin V indicate that there are no differences between elutriated fractions immediately upon purification or after freezing and thawing. The studies also indicate that interdonor variations are much larger than intradonor variations. CONCLUSIONS Although it differs from other reports in certain respects, our procedure has nevertheless produced results in line with other findings. After extensive testing and use in different contexts we feel confident that we have established a method for producing a large number of purified and well-preserved monocytes. CONCLUSIONS The goal of being able to perform a large number of experiments with monocytes of high purity and good functionality has been reached.

UI MeSH Term Description Entries
D009000 Monocytes Large, phagocytic mononuclear leukocytes produced in the vertebrate BONE MARROW and released into the BLOOD; contain a large, oval or somewhat indented nucleus surrounded by voluminous cytoplasm and numerous organelles. Monocyte
D001772 Blood Cell Count The number of LEUKOCYTES and ERYTHROCYTES per unit volume in a sample of venous BLOOD. A complete blood count (CBC) also includes measurement of the HEMOGLOBIN; HEMATOCRIT; and ERYTHROCYTE INDICES. Blood Cell Number,Blood Count, Complete,Blood Cell Counts,Blood Cell Numbers,Blood Counts, Complete,Complete Blood Count,Complete Blood Counts,Count, Blood Cell,Count, Complete Blood,Counts, Blood Cell,Counts, Complete Blood,Number, Blood Cell,Numbers, Blood Cell
D002469 Cell Separation Techniques for separating distinct populations of cells. Cell Isolation,Cell Segregation,Isolation, Cell,Cell Isolations,Cell Segregations,Cell Separations,Isolations, Cell,Segregation, Cell,Segregations, Cell,Separation, Cell,Separations, Cell
D002499 Centrifugation, Density Gradient Separation of particles according to density by employing a gradient of varying densities. At equilibrium each particle settles in the gradient at a point equal to its density. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Centrifugations, Density Gradient,Density Gradient Centrifugation,Density Gradient Centrifugations,Gradient Centrifugation, Density,Gradient Centrifugations, Density
D005434 Flow Cytometry Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake. Cytofluorometry, Flow,Cytometry, Flow,Flow Microfluorimetry,Fluorescence-Activated Cell Sorting,Microfluorometry, Flow,Cell Sorting, Fluorescence-Activated,Cell Sortings, Fluorescence-Activated,Cytofluorometries, Flow,Cytometries, Flow,Flow Cytofluorometries,Flow Cytofluorometry,Flow Cytometries,Flow Microfluorometries,Flow Microfluorometry,Fluorescence Activated Cell Sorting,Fluorescence-Activated Cell Sortings,Microfluorimetry, Flow,Microfluorometries, Flow,Sorting, Fluorescence-Activated Cell,Sortings, Fluorescence-Activated Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D013997 Time Factors Elements of limited time intervals, contributing to particular results or situations. Time Series,Factor, Time,Time Factor
D015925 Cryopreservation Preservation of cells, tissues, organs, or embryos by freezing. In histological preparations, cryopreservation or cryofixation is used to maintain the existing form, structure, and chemical composition of all the constituent elements of the specimens. Cryofixation,Cryonic Suspension,Cryonic Suspensions,Suspension, Cryonic
D017304 Annexin A5 A protein of the annexin family isolated from human PLACENTA and other tissues. It inhibits cytosolic PHOSPHOLIPASE A2, and displays anticoagulant activity. Annexin V,Placental Anticoagulant Protein I,Anchorin CII,Calphobindin I,Endonexin II,Lipocortin V,Lipocortin-V
D018950 Lipopolysaccharide Receptors Glycolipid-anchored membrane glycoproteins expressed on cells of the myelomonocyte lineage including MONOCYTES; MACROPHAGES; and some GRANULOCYTES. They function as receptors for the complex of lipopolysaccharide (LPS) and LPS-binding protein. Antigens, CD14,CD14 Antigens,Receptors, Lipopolysaccharide,Soluble CD14,Soluble CD14 Antigen,Soluble CD14 Protein,sCD14,CD14 Antigen,CD14 Monocyte Differentiation Antigen,LPS Receptor,Lipoglycan Receptor,Receptor, LPS,Receptor, Lipoglycan,CD14 Antigen, Soluble,CD14 Protein, Soluble,CD14, Soluble

Related Publications

P K Lund, and G B Joø, and A B Westvik, and R Ovstebø, and P Kierulf
Isolation of monocytes from whole blood-derived buffy coats by continuous counter-flow elutriation.
October 2006, Journal of clinical apheresis,
P K Lund, and G B Joø, and A B Westvik, and R Ovstebø, and P Kierulf
Isolation of human monocytes by Ficoll density gradient centrifugation.
December 1981, Blut,
P K Lund, and G B Joø, and A B Westvik, and R Ovstebø, and P Kierulf
Isolation of functionally different human monocytes by counterflow centrifugation elutriation.
July 1982, Blood,
P K Lund, and G B Joø, and A B Westvik, and R Ovstebø, and P Kierulf
Isolation of ovine lymphocytes, granulocytes, and monocytes by counterflow centrifugation elutriation.
August 1982, American journal of veterinary research,
P K Lund, and G B Joø, and A B Westvik, and R Ovstebø, and P Kierulf
Isolation of leukemic blasts from peripheral blood by density gradient centrifugation.
April 1970, The Journal of laboratory and clinical medicine,
P K Lund, and G B Joø, and A B Westvik, and R Ovstebø, and P Kierulf
Isolation of megakaryocytes by a combination of density gradient centrifugation and velocity sedimentation, and centrifugal elutriation.
December 1985, The Kobe journal of medical sciences,
P K Lund, and G B Joø, and A B Westvik, and R Ovstebø, and P Kierulf
Isolation of human megakaryocytes by density centrifugation and counterflow centrigual elutriation.
May 1984, The Journal of laboratory and clinical medicine,
P K Lund, and G B Joø, and A B Westvik, and R Ovstebø, and P Kierulf
A system for obtaining large numbers of cryopreserved human monocytes purified by leukapheresis and counter-current centrifugation elutriation (CCE).
September 1983, Journal of immunological methods,
P K Lund, and G B Joø, and A B Westvik, and R Ovstebø, and P Kierulf
Purification of human blood monocytes by hypotonic density gradient centrifugation in Percoll.
November 1982, Journal of immunological methods,
P K Lund, and G B Joø, and A B Westvik, and R Ovstebø, and P Kierulf
Separation of Crassostrea gigas hemocytes by density gradient centrifugation and counterflow centrifugal elutriation.
January 1988, Developmental and comparative immunology,
Copied contents to your clipboard!