Method for separating lymphocytes

Abstract

The invention discloses a method for separating lymphocytes and belongs to the technical field of biology. The method comprises the following steps: fully dissolving the following components that meet the medical standards: 3 to 5 parts by weight of dextran, 2 to 4 parts by weight of mycose, 0.3 to 0.8 part by weight of sodium alginate with an ultralow viscosity, 1 to 3 parts by weight of meglumine diatrizoate, and 0.1 to 0.15 part by weight of poly(diallyl dimethyl ammonium chloride) into sterile water to prepare 100 parts by weight of physiological solution; filtering the physiological solution by a filter membrane to remove the bacteria to obtain a separated liquid; adding the separated liquid into a centrifuge tube, wherein the volume of the separated liquid is not more than 1 / 2 of the volume of the centrifuge tube; paving a sample, which has been processed by an anticoagulant and diluted by a balanced salt solution or serum-free culture medium, on the separated liquid, carrying out centrifugation for 10 to 30 minutes at a temperature of 8 to 25 DEG C under a gravity of 200 to 800 g; wherein the sample is derived from peripheral blood, umbilical cord blood or marrow. The obtained lymphocytes have a good tumor killing activity.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a separation method for separating lymphocytes. Background technique [0002] During the standard separation process, the cells must be in contact with the separation medium for 15-45 minutes to achieve the best separation effect. During this period, the components in the separation solution may accumulate inside the cells through passive diffusion, active cell transport, endocytosis / pinocytosis, surface adhesion, etc., and this part of the accumulated components is difficult to pass through conventional methods Washing completely removes the isolated target cells (eg, lymphocytes). [0003] Moreover, the current gradient centrifugation method has cumbersome steps, strong skills, and relatively high requirements for the operator's experience: gradient centrifugation requires three steps, especially the second step. Density of the cell suspension without confusing the interface will...

AI Technical Summary

Problems solved by technology

[0003] Moreover, the current gradient centrifugation method has cumbersome steps, strong skills, and relatively high requirements for the operator's experience: gradient centrifugation requires three steps, especially the second step. Density of the cell suspension without mixing the interface, which greatly tests the operator's experience and patience, time-consuming, laborious, and prone to fatigue, and once the liquid level is mixed, the separation effect will be greatly affected, and even the lymphocyte layer cannot be separated

Moreover, most of the current separation fluids have certain toxicity, which affects the separation effect and the quality of the separated cells, resulting in low separation efficiency.