Method for fast and efficient separation and purification of tumor cells from malignant pleural effusion and ascites

Abstract

The invention discloses a method for fast and efficient separation and purification of tumor cells from malignant pleural effusion and ascites. The method includes the steps of: (1) conducting aseptic operation to collect malignant pleural effusion and ascites, conducting subpackaging in sterile centrifuge tubes, and performing centrifugation at room temperature to obtain cell precipitate; (2) conducting re-suspension on the cell precipitate with an aseptic PBS liquid to obtain a tumor cell suspension, adding the tumor cell suspension onto the serum in the centrifuge tubes along the pipe walls of the centrifuge tubes to form a two-layer solution in the centrifuge tubes, with the upper layer being a tumor cell suspension layer and the lower layer being a serum layer; and (3) standing the centrifuge tubes added with the tumor cell suspension in step (2) for 5-8min, then sucking the serum at the tube bottoms by a pasteur pipet without destroying liquid layering, placing the sucked serum in a new centrifuge tube, adding aseptic PBS liquid, conducting centrifugation at room temperature, and discarding the supernatant, thus obtaining purified tumor cells. The method provided by the invention is suitable for separation and purification of tumor cells in various malignant pleural effusion and ascites, and avoids the limitations of existing separation and purification methods.

Description

technical field [0001] The invention relates to a method for separating tumor cells, in particular to a method for rapidly and efficiently separating and purifying tumor cells from malignant pleural ascites. Background technique [0002] Pleural effusion is a common clinical symptom in respiratory medicine. It is divided into two types: exudative and exudative. It can be caused by inflammatory lesions, neoplastic lesions, systemic diseases, and certain organ lesions. Among them, malignant pleural effusion is mainly caused by malignant tumors of thoracic organs, especially common in advanced lung cancer, with an incidence rate as high as 60%. And often progressive. The main cause of the disease is the invasion and obstruction of capillaries and lymphatic vessels by pleural metastatic nodules. Therefore, the pleural effusion contains a large amount of protein and formed blood components, and bloody pleural effusion accounts for about 75%. [0003] Ascites is excessive fluid ...

AI Technical Summary

Problems solved by technology

However, these methods all have different defects, such as the small amount of cell collection, separation and purification steps, low degree of purification, and the need for special separation and purification reagents such as tumor cell separation liquid and lymphocyte separation liquid.

Moreover, the operation of the above method involves multiple times of long-time high-speed centrifugation, which may easily lead to poor cell status after separation, and the cells do not adhere to the wall, making further research impossible.

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