| 摘要 |
FIELD: biotechnology.SUBSTANCE: viable lymphocytes are obtained from the walls of the small intestine (lamina propria) of CBA mice while using biological material of three mice. The source biological material is a part of intestine, which is washed with phosphate-buffered solution, and the Peyer's glands are removed. It is milled in the presence of phosphate-saline solution containing dithiothreitol under stirring at room temperature. It is washed with the buffer solution through a nylon mesh. The material is incubated in the buffered solution containing bovine serum albumin with stirring under the specified conditions. The supernatant liquor is removed. The precipitate is washed with phosphate-saline buffer. The precipitate of the biological material is incubated in the cellular medium RPMI-1640 comprising fetal calf serum, HEPES and collagenase with stirring under the specified conditions, and the resulting suspension is filtered. The suspension is washed from the residues of collagenase, and the lymphocyte cells are precipitated in the cellular medium RPMI-1640 comprising fetal calf serum and HEPES using centrifugation under specified conditions, followed by determination of yield of viable cells using the Goryaev chamber.EFFECT: invention enables to increase the yield of viable cells from the small intestine of a mouse.1 ex |
