Cell line for lowly-metastatic clear cell carcinoma of kidney of Han Chinese

A renal clear cell carcinoma and clear cell technology, applied in the field of microbial animal cell lines, can solve problems such as the lack of systematic and in-depth forward-looking basic theoretical research

Inactive Publication Date: 2009-10-07
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Abstract
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  • Application Information

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Problems solved by technology

At present, research on renal cell carcinoma mainly focuses on the improvement of diagnostic methods, selection of treatment options, and retrospective pathological analysis, lacking systematic and in-depth prospective basic theoretical research

Method used

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  • Cell line for lowly-metastatic clear cell carcinoma of kidney of Han Chinese
  • Cell line for lowly-metastatic clear cell carcinoma of kidney of Han Chinese
  • Cell line for lowly-metastatic clear cell carcinoma of kidney of Han Chinese

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Obtainment of the low-metastatic clear cell renal cell carcinoma cell line LoMet-ccRCC of the Han nationality

[0037] The LoMet-ccRCC cells of the present invention are obtained from the kidney tumor tissue in situ of a Han patient with renal clear cell carcinoma. Put the resected tumor tissue into a glass plate filled with 5ml D-Hanks solution, and remove necrotic tissue, adipose tissue, blood vessels, etc. Soak the tissue block in 5-10ml 1640 medium for 30-40min, cut the tissue block into 1-3mm 3 For small pieces, transfer the soaking liquid and tissue pieces to a centrifuge tube, shake and wash repeatedly for 2-3 minutes, centrifuge at 1500 rpm for 10 minutes, discard the supernatant, add 5-10ml of 1640 medium to suspend the tissue pieces, shake and wash repeatedly 2-3min, 1500 rpm, centrifuge for 10min, discard the supernatant, suspend the tissue pieces with 3-4ml of 1640 complete medium, and transfer the suspension to a cell culture bottle at 1ml / bottle...

Embodiment 2

[0038] Example 2 LoMet-ccRCC cell growth curve

[0039] will be 3×10 4 Inoculate cells into 24-well plates, add 2ml of 1640 complete medium, and inoculate 21 wells in total. Thereafter, 3 wells were digested every other day and cell counts were performed. Calculate the mean and standard deviation of the cell number, and draw the cell growth curve. Such as figure 2 As shown, the growth of LoMet-ccRCC cells was relatively stable, and entered exponential growth on the 3rd day after inoculation, reached the fastest growth rate on the 4th day, and then gradually slowed down. The cell doubling time was 16.5h.

Embodiment 3

[0040] Example 3 Chromosome analysis of LoMet-ccRCC cells

[0041] The number of chromosomes in 100 metaphase cells was counted, and the number of chromosomes in LoMet-ccRCC cell lines was mainly concentrated between 50 and 70, and the mode of chromosomes was 55. Prompt the presence of hyperdiploidy and so on.

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Abstract

The invention discloses a cell line for lowly-metastatic clear cell carcinoma of kidney of Han Chinese, namely LoMeT-ccRcc CCTCC No.C200910. The cell line can grow for a long time in vitro and be stably handed down, has epithelioid cell form, and has no contact inhibition. The cell is heteroploid, chromosome number and structure are aberrant. Immunohistochemistry shows masculine CA9 and CD133. A flow cytometry analyses and finds out that CA9 and CD133 expression is obviously enhanced in the condition that cell is handed down for many times. The cell line has lowly-metastatic trend after the cell is handed down continuously. The recovery rate of the cell line after being frozen is more than 80%, the recovered cell growth state accords with the original cell. The cell line of the invention is used for establishing a cell model for sieving and preparing medicines that is used for dialogising, preventing and treating clear cell carcinoma of kidney.

Description

technical field [0001] The invention relates to a microbial animal cell line, in particular to a Han Chinese low-metastatic renal clear cell carcinoma cell line LoMet-ccRCC, CCTCC-C200910. Background technique [0002] Tumor cell lines play an important role in the basic research of tumors. The use of cell lines for in vitro experiments and the construction of in vivo animal models has become a research hotspot, and the research results obtained from them are comparable and convincing. At present, the in vitro culture of human cancer cells has become a widely used method in medical and biological research. It can not only study the mechanism of tumor occurrence and development at the molecular and gene levels, but also has great significance for clinical diagnosis, drug screening and tumor treatment. [0003] Renal cell carcinoma is the most common solid malignant tumor of the kidney (accounting for about 96%), ranking second among genitourinary system tumors, accounting for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12Q1/18C12R1/91
Inventor 曹广文翟羽佳苏彤贺松琴常文军
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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