Method for establishing fluorescent orthotopic transplantation model for subcutaneous transferred organ cancer

A fluorescence in situ and model building technology, applied in the field of visceral organ cancer research, can solve the problem that observation is not very easy

Inactive Publication Date: 2009-08-12
NANJING ORIGIN BIOSCI
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But if the tumor is growing in the abdominal cavity, e

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  • Method for establishing fluorescent orthotopic transplantation model for subcutaneous transferred organ cancer
  • Method for establishing fluorescent orthotopic transplantation model for subcutaneous transferred organ cancer

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Embodiment Construction

[0010] Materials and Methods:

[0011] 1. Experimental animals: BALB / C nu / nu nude mice, SPF grade, purchased from Beijing Kelihua Experimental Animal Center, weighing 24-28g, were kept in separate air-supplied cages, and the room temperature was controlled at 24°C-25°C. The relative humidity is 50% to 60%.

[0012] 2. Cell lines and cell culture: select the cell lines from the corresponding tumors, culture in RPMI-1640 medium, 10% calf serum at 37°C, 5% CO2, digest with 0.25% trypsin-EDTA, wash twice with PBS, Adjust cell concentration to make 1 x 10 6 cell suspension.

[0013] 3. Construction of green fluorescent protein (GFP) pLPCX retroviral plasmid and transfection of tumor cells: the expression plasmid pEGFP-N3(-) containing GFP was purchased from BD Biosciences Clontech (BD Biosciences, CAUSA), and the retrovirus pLPCX was purchased from From Clontech Laboratories, Inc. (Takara, Japan). Primers containing restriction endonuclease Bgl II and Cla I sites were used to am...

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Abstract

The invention relates to a method for establishing a model of fluorescence orthotopic transplantation of subcutaneous metastasis organ cancers, which is characterized in that human colon adenocarcinoma cell strains or cancer cell strain green fluorescent proteins (GFP) of corresponding organs are selected; GFP plasmids carry out transfection on cancer cell strains of the organs via retrovirus pLPCX so that the cancer cell strains of the organs contain GFP expression plasmids; a GFP gene is connected with a pLPCX vector via a T4 ligase; a GFP-pLPCX vector is transfected to a PT67 package cell to obtain a Lovo cell; nude mice are selected and subjected to subcutaneous inoculation with Lovo cell suspension; tumors can be formed in about 4 weeks; orthotopic transplantation is carried out after green fluorescence is observed; a cut is opened on the skin and peritoneum; about 1cm cecum of the nude mice is taken out of the peritoneum; the gap between the cecum and the peritoneum is sutured and the cecum is transferred subcutaneously; the cut cecum is subjected to serosa; two fluorescence-carrying tumors with diameter of about 1mm are transplanted to the cecum via the suture and the skin is closed.

Description

1. Technical field [0001] The invention relates to a method for studying canceration of visceral organs (such as colon and other organs), in particular to a method for establishing an orthotopic transplantation model of colon canceration. 2. Background technology [0002] Chinese patent CN200710019273.0 is a method for establishing a colorectal cancer stoma orthotopic transplantation model invented by the present application, which includes the following steps: a) cell line selection or cell culture, which is prepared by using colon adenocarcinoma cell line cell concentration ( 1±0.5)×10 6 b) the colostomy in living animals, the steps are to put the cecum out of the abdominal wall, depending on the length of the experimental animal, it is 0.3-2cm and fix and heal with the surrounding skin; d) orthotopic transplantation of the tumor at the stoma, The steps are as follows: take 0.1-1ml of mouse colon adenocarcinoma cell suspension and inoculate it under the stoma mucosa to gr...

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Application Information

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IPC IPC(8): A61B19/00G01N21/64A61B90/00
Inventor 金黑鹰杨志坚
Owner NANJING ORIGIN BIOSCI
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