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Method for establishing mouse transplanting tumor model with normal immunologic function

A method of establishing a method and a technology of immune function, which is applied in the field of transplanted tumor models in mice, and can solve problems other than tumor immunotherapy and immune function deficiency

Inactive Publication Date: 2011-07-20
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the lack of T cells in nude mice, SCID mice lack T cells and B cells at the same time, they have immune function defects, and are not the most ideal model for studying tumor immunotherapy

Method used

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  • Method for establishing mouse transplanting tumor model with normal immunologic function
  • Method for establishing mouse transplanting tumor model with normal immunologic function

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Experimental program
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Embodiment 1

[0018] The vector expressing Ras mutant gene (see Serrano M, Lin AW, McCurrach ME, Beach D, Lowe SW (1997). Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88: 593-602.) and expression The carrier of the p53 mutant gene (p53S) (see another patent application of the applicant: the establishment method in the patent acceptance number 200910095184.3: the cDNA of the p53 mutant gene is connected into the PQCXIP carrier by conventional genetic engineering methods, so that p53 can be expressed mutant gene) transfected into p53 - / - Mouse embryonic fibroblasts (MEFs) were screened for p53 - / - +S+Ras cell line; transfect the vector carrying the mutant Ras gene and the empty plasmid into p53 - / - MEF cells get p53 - / - +V+Ras cell line.

[0019] 1) p53 - / - +V+Ras and p53 - / - +S+Ras cells were subcultured in vitro by adding DMEM (10% FBS).

[0020] 2) Take 6 dishes of each of the two types of cells, suck off the supernatant, and...

Embodiment 2

[0028] p53 - / - +V+Ras and p53 - / - +S+Ras cells (see the first part of the embodiment: the vector carrying the Ras gene and the empty plasmid transfected into p53 - / - MEF cells get p53 - / - +V+Ras cell line. ) into DMEM (10% FBS) for subculture in vitro; take 6 dishes of each of the two types of cells, suck off the supernatant, add 5ml of PBS to wash once; after trypsinization, add 4ml of medium, collect the cells into a centrifuge tube; centrifuge at 1200rpm for 5min , discard the supernatant; add 7ml DMEM (serum-free), resuspend, take 100μl for cell counting; use DMEM (serum-free) to make 2×10 6 / ml two-cell suspension, take the two-cell suspension and inoculate it subcutaneously on the back of different C57 / BL6 mice. 6 cells. On average, tumors can be observed about 14 to 5 days after inoculation of the two kinds of cells, and an allogenic mouse transplanted tumor model with normal immune function can be obtained.

Embodiment 3

[0030] p53 - / - +V+Ras and p53 - / - +S+Ras cells (see the first part of the embodiment: the vector carrying the Ras gene and the empty plasmid transfected into p53 - / - MEF cells get p53 - / -+V+Ras cell line. ) into DMEM (10% FBS) for subculture in vitro; take 6 dishes of each of the two types of cells, suck off the supernatant, add 5ml of PBS to wash once; after trypsinization, add 4ml of medium, collect the cells into a centrifuge tube; centrifuge at 1200rpm for 5min , discard the supernatant; add 7ml DMEM (serum-free), resuspend, take 100μl for cell counting; use DMEM (serum-free) to make 0.6×10 6 / ml two-cell suspension, take the two-cell suspension and inoculate it subcutaneously on the back of different C57 / BL6 mice. 6 cells. On average, tumors can be observed about 14 to 5 days after inoculation of the two kinds of cells, and an allogenic mouse transplanted tumor model with normal immune function can be obtained.

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Abstract

The invention relates to a method for establishing a mouse transplanting tumor model with a normal immunologic function. The mouse transplanting tumor model with the normal immunologic function is successfully established by utilizing a wild mouse as a carrier and utilizing p53- / -+S+Ras and p53- / -+V+Ras mouse tumor cells which are near-source, have clear molecular genetic background and carry conventional p53 and Ras genic mutation in the human tumor. Compared with the traditional mouse transplanting tumor model with immunodeficiency, the model provided by the invention more faithfully simulates the condition of the human tumor disease and can be used for evaluating a medicament for achieving the tumor resistant effect by improving the body immunologic function. The mouse tumor model established by the method has the advantages of short period and high repeatability. In addition, by using the mouse tumor cell with clear molecular genetic background, the pharmacological action molecular mechanism research in animals can be carried out.

Description

Technical field: [0001] The invention belongs to the field of medicine, and in particular relates to a method for establishing a mouse transplanted tumor model with normal immune function, and a mouse transplanted tumor model with normal immune function obtained by the method, and their application in the field of pharmacy in the application. Background technique [0002] Tumor is a kind of frequently-occurring and common disease that seriously threatens human health. Its pathogenesis has not been fully elucidated, and the treatment effect is not satisfactory. Tumors are genetic diseases in nature, caused by various environmental and genetic carcinogens in a synergistic or sequential manner. The TP53 gene is the gene most closely related to human tumor suppression found so far. In just over a decade, people's understanding of the TP53 gene has undergone three transformations from oncoprotein antigens, oncogenes to tumor suppressor genes. Under normal circumstances, the exp...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K49/00C12N5/10
Inventor 罗瑛唐文如赵兰君
Owner KUNMING UNIV OF SCI & TECH
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