FIELD: medicine.
SUBSTANCE: method involves bone marrow sample obtained by aspiration in puncturing iliac bone crest. It is layered over density gradient of 1.077 g/l and centrifuged 30 min long at 400 g and temperature of 18-22°C. Mononuclear cells are collected from phase separation boundary. To take 100 ml of bone marrow by aspiration, 20 ml of heparinized physiologic saline is introduced with heparin concentration of 2500 units/ml. Before layering, the heparinized aspirated bone marrow IS preliminarily precipitated during 30 min at temperature of 18-22°C. Then, upper 2/3 of cellular suspension is taken and the sample is diluted twice as much with heparinized phosphate-bufferized physiologic saline having heparin concentration of 10 units/ml. After having finished layering and centrifuging, the cell layer, collected from phase separation boundary, is washed with heparinized phosphate-bufferized physiologic saline having heparin concentration of 10 units/ml by means of triple centrifuging 10 min long each, at 400 g and temperature of 18-22°C, and the cellular suspension is brought to required concentration.
EFFECT: higher amount of viable cells.
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Authors
Dates
2007-02-10—Published
2005-04-18—Filed