Method for simultaneous separation of T and B lymphocytes in peripheral blood

Abstract

The invention discloses a method for simultaneous separation of T and B lymphocytes in peripheral blood. The method is an improved nylon wool column separation method, can realize one-time simultaneous separation, so that two kinds of lymphocytes, namely T and B lymphocytes are obtained, cell yield is high, cross contamination is less, there is no effect on the biological activity of the cells, and the next step of the experimental study is easy to carry out.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and relates to a high-efficiency separation and culture method for simultaneously obtaining peripheral blood T and B lymphocytes. Background technique [0002] Isolation of lymphocytes from peripheral blood is an important technique in modern molecular biology and immunology research. In recent years, cell separation technology has made great progress. At present, there are four main lymphocyte separation technologies: nylon hair column method, immunomagnetic bead method, gradient centrifugation method, and flow sorting method. The relative sorting cost of flow cytometry is relatively high, requiring large-scale flow sorting instruments, and the operation is laborious. At the same time, the sorting and screening process of flow cytometry may sometimes affect the activity and proliferation of cells. Although the purity of the cells obtained by the immunomagnetic bead method is high, the cell...

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Problems solved by technology

The relative sorting cost of flow cytometry is relatively high, and large-scale flow sorting instruments are required, which is laborious to operate. At the same time, the sorting and screening process of flow cytometry may sometimes affect the activity and proliferation of cells

Although the purity of the cells obtained by the immunomagnetic bead method is high, the cell acquisition rate is very low, and the lymphocytes are easily activated by the immunomagnetic bead positive sorting antibody to activate downstream signaling pathways, which affects subsequent research

Although the gradient centrifugation method obtains a large number of cells, the purity of the cells is poor

The yield of lymphocytes obtained by the traditional nylon hair column method is relatively low, and the interaction between T and B lymphocytes cannot be distinguished

Moreover, for the separation of lymphocytes in peripheral blood, the usual separation technology has low cell separation efficiency, and its activity is easily affected by magnetic bead antibodies and lost antibodies, and it cannot effectively separate and distinguish T lymphocytes and B lymphocytes, which is likely to cause T and B lymphocytes. Interaction and cross-contamination between two types of lymphocytes

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