Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Method of detaching cells – digesting tissue or establishing...
Reexamination Certificate
2006-12-28
2010-06-01
Davis, Ruth A (Department: 1651)
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
Method of detaching cells, digesting tissue or establishing...
C435S372000, C435S379000
Reexamination Certificate
active
07727764
ABSTRACT:
A method of “non-isopycnic” cell isolation and purification has the steps of adding a sample of blood to a defined volume to a corresponding ratio volume of EDTA solution to produce a volume of anti-coagulated blood; taking a predetermined volume of the anti-coagulated blood and placing it in a first tube containing a selected defined volume of PSS, wherein the selected defined volume of PSS is taken from a group of defined volumes of PSS, as each defined volume of PSS establishes a specific cell type to be purified; centrifuging the tube for a first predetermined time and speed to form a volume of supernatant of plasma/PSS and a bottom sedimented volume of mostly red blood cells; extracting the supernatant to within a proximity of an interface between the sedimented red blood cells and the supematant; transferring an appropriate, pre-selected volume amount of the supernatant into a second tube holding a defined volume of physiological media; mixing the solutions gently; centrifuging for a second predetermined speed and time; and pouring off the supernatant wherein at the bottom of the tube will be a cell button containing a volume of the selected purified cells in a high percentage and a very small quantity or low percentage of some contaminating non-selected cells. This method is useful in establishing high purity concentrations of selected cells from whole blood such as monocytes, lymphocytes, neutrophils and basophils.
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Percoll Product Information, Pharmacia, Sigma Aldrich, Inc.
Raghuprasad, Puthalath K.; A Rapid Simple Method Of Basophil Purification By Density Centrifugation On Percoll; The Journal Of Immunology; Nov. 1982; vol. 129, No. 5; pp. 2128-2133; The American Association of Immunologists, USA.
Davis Ruth A
King David L.
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