Kidney cancer circulating tumor cell line and acquisition, separation and culture method

Abstract

The invention provides a kidney cancer circulating tumor cell line and an acquisition, separation and culture method. The method comprises the following steps: S1, establishing a stably-transfected human kidney cancer cell line 786-O, infecting the human kidney cancer cell line 786-O with viruses to stably transfect the human kidney cancer cell line 786-O into green fluorescent protein and luciferase, enabling the human kidney cancer cell line 786-O to have Puromycin resistance, and culturing the human kidney cancer cell line 786-O; S2, preparing a circulating tumor cell animal model, injecting the human kidney cancer cell 786-O cultured in the step S1 into a mouse body through caudal vein injection, feeding the mouse in an SPF environment for one month, and observing whether metastasis occurs or not through small animal imaging; S3, extracting a cell line of circulating tumor cells, collecting peripheral blood of the mouse subjected to metastasis in the step S2, separating, and culturing to obtain kidney cancer circulating tumor cells; and S4, culturing the circulating tumor cells obtained in the step S3. The method is simple and easy to operate, high in success rate and low in cost, the obtained circulating tumor cells are extremely high in purity and good in cell state, and the method can be widely applied to clinical research.

Description

Technical field[0001]The invention belongs to the technical field of separation and culture of tumor cells, and specifically relates to a kidney cancer circulating tumor cell line and methods for obtaining, separating and culturing.Background technique[0002]Circulating tumor cells, as tumor cells that spread into the circulating blood, play an extremely important role in the development of cancer. In the early stage of the disease, circulating tumor cells undergo epithelial mesenchysis to gain invasiveness, and then enter the circulating blood with the help of immune cells, and enter a dormant state after reaching remote organs. As the tumor progresses, it will eventually recover and metastasize under certain conditions. The capture technology for circulating tumor cells has also evolved from the initial antibody capture to a new technology that captures the physical properties of circulating tumor cells and integrates the two capture methods. The emergence of new technologies has i...

AI Technical Summary

Problems solved by technology

The circulating tumor cells isolated by this method can be normally subcultured and can be applied to clinical research. However, in the preparation of circulating tumor cell animal models and the extraction of circulating tumor cell cell lines, the survival rate of mice is still low and the success rate of modeling is low. , high economic cost, complex operation steps and other defects

Images