Isolation and culture method of mouse renal clear cell carcinoma circulating tumor cell line and human-derived clear cell renal carcinoma circulating tumor cell

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

Active Publication Date: 2022-05-06
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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  • Isolation and culture method of mouse renal clear cell carcinoma circulating tumor cell line and human-derived clear cell renal carcinoma circulating tumor cell
  • Isolation and culture method of mouse renal clear cell carcinoma circulating tumor cell line and human-derived clear cell renal carcinoma circulating tumor cell
  • Isolation and culture method of mouse renal clear cell carcinoma circulating tumor cell line and human-derived clear cell renal carcinoma circulating tumor cell

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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 method for separating and cultivating circulating tumor cell lines of clear cell renal cell carcinoma of mice and circulating tumor cells of clear cell renal cell carcinoma, and relates to the technical field of separation and culturing of tumor cells; the circulating tumor of clear cell renal cell carcinoma of mice The cell line is circulating tumor cells of orthotopic renal carcinoma NOD / SCID mice; the method for isolating and culturing the circulating tumor cells mainly includes the following steps: first, human-derived clear cell renal cell carcinoma cell line 786-O is infected with virus, and the It stably transforms green fluorescent protein and has Puromycin resistance, and cultures the cell line; then establishes a mouse model, collects the peripheral blood of the mouse, cultures it, and separates it to obtain circulating tumor cells of renal cancer. The separation and culture method of the present invention is simple and easy to operate, and the isolated circulating tumor cells can be cultured in normal leaflets, and can be widely used in 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 Patents(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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