Separation and culture method of mouse kidney clear cell carcinoma circulating tumor cell line and human kidney clear cell carcinoma circulating tumor cells

A technology for renal clear cell carcinoma and tumor cells, applied in the direction of cell culture active agents, biochemical equipment and methods, tissue culture, etc., can solve problems such as separation and culture methods for circulating tumor cells of renal cancer that have not been seen yet, and achieve simple culture methods Easy to operate, strong proliferation, high purity effect

Active Publication Date: 2020-06-23
SECOND AFFILIATED HOSPITAL OF COLLEGE OF MEDICINEOF XIAN JIAOTONG UNIV
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Problems solved by technology

So far, there have been no reports on the isolation and culture of circulating tumor cells from renal cell carcinoma

Method used

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  • Separation and culture method of mouse kidney clear cell carcinoma circulating tumor cell line and human kidney clear cell carcinoma circulating tumor cells
  • Separation and culture method of mouse kidney clear cell carcinoma circulating tumor cell line and human kidney clear cell carcinoma circulating tumor cells
  • Separation and culture method of mouse kidney clear cell carcinoma circulating tumor cell line and human kidney clear cell carcinoma circulating tumor cells

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Embodiment Construction

[0031] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0032] The present invention is a mouse renal clear cell carcinoma circulating tumor cell line of an embodiment, which is a circulating tumor cell of orthotopic renal carcinoma NOD / SCID mouse

[0033] The method for separating and culturing circulating tumor cells of renal clear cell carcinoma according to the embodiment of the present invention is as follows:

[0034] (1) Reagents used in the experiment

[0035] Lentivirus EF1A-MCS-P2A-EGFR-SV40-firefly_Luc-IRES-Puromycin (Gibco Gene); Polybrene dye aid (Gibco Gene); red blood cell lysate (Biyuntian); epithelial growth factor EGF (Gibco); alkali ...

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Abstract

The invention discloses a separation and culture method of a mouse kidney clear cell carcinoma circulating tumor cell line and kidney clear cell carcinoma circulating tumor cells, and relates to the technical field of separation and culture of tumor cells. The mouse kidney clear cell carcinoma circulating tumor cell line is circulating tumor cells of an in-situ kidney cancer NOD/SCID mouse. The separation and culture method of the circulating tumor cells mainly comprises the following steps of: firstly, infecting a human kidney clear cell carcinoma cell line 786-O with viruses, so that the human kidney clear cell carcinoma cell line is stably converted into green fluorescent protein and has Puromycin resistance, and culturing the cell line; and then establishing a mouse model, collecting peripheral blood of the mouse, and culturing and separating the peripheral blood to obtain the kidney cancer circulating tumor cells. The separation and culture method disclosed by the invention is simple and easy to operate, and the circulating tumor cells obtained by separation can be subjected to normal leaflet culture and can be widely applied to clinical research.

Description

technical field [0001] The invention relates to the technical field of separation and cultivation of tumor cells, in particular to a method for the separation and cultivation of a mouse renal clear cell carcinoma circulating tumor cell line and human-derived renal clear cell carcinoma circulating tumor cells. Background technique [0002] Renal cell carcinoma (RCC) is the most common malignant tumor in the kidney. 20%-30% of patients will relapse after treatment, and there is a high possibility of death from the tumor. Malignant tumor cells can fall off from the primary tumor and enter the blood circulation in the early stage of cancer, and use the circulatory system to reach distant organs and enter a dormant state or form metastases. Circulating tumor cells (CTCs) are disseminated tumor cells in the peripheral blood of cancer patients, which shed from primary tumors or metastatic sites into peripheral blood, and can be used as liquid biopsies for early diagnosis, risk asse...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2501/11C12N2501/115
Inventor 王振龙李刚管栩冰种铁
Owner SECOND AFFILIATED HOSPITAL OF COLLEGE OF MEDICINEOF XIAN JIAOTONG UNIV
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