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Green fluorescent human liver cancer cell line capable of transfer

A human liver cancer cell line and green fluorescence technology, applied in the field of human liver cancer cell lines, can solve problems such as difficulty in effect, inability to observe tumor growth in real time, angiogenesis of recurrent and metastatic tumors, inability to early detect liver cancer recurrence, and tracking of metastases and metastases

Inactive Publication Date: 2004-11-17
ZHONGSHAN HOSPITAL FUDAN UNIV
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Problems solved by technology

However, looking at the human liver cancer models at home and abroad, it is impossible to detect the recurrence and metastasis of liver cancer early and trace the metastases, and it is also impossible to observe the growth, recurrence and metastasis of the tumor in real time, and the tumor angiogenesis closely related to it. Difficulty assessing the effects of medications or other interventions

Method used

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  • Green fluorescent human liver cancer cell line capable of transfer
  • Green fluorescent human liver cancer cell line capable of transfer
  • Green fluorescent human liver cancer cell line capable of transfer

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Establishment of human liver cancer cell lines with high green fluorescence and low metastases.

[0022] 1. MHCC97-H and MHCC97-L cells were divided into 3×10 5 Add cells / well to a six-well plate, add 2ml DMEM medium (GIBCOBRL, Grand Island, NY, USA) containing 10% FCS (Hyclone, Utah, USA) to each well, and incubate until the cells are 60-80% confluent;

[0023] 2. Replace 1ml of DMEM (plus 5% FCS) medium on the day of transfection, and incubate at 37°C for 1 hour;

[0024] 3. Prepare transfection solution: solution A: 1.5ug of pIRES2-EGFP (PT3267-5, ClontechLaboratories Inc., Palo Alto, CA, USA) plasmid DNA was diluted to 50ul (30ug / ml) / well with HBS buffer; Solution B: 6ul DOSPER (Cat.No.1811169, Roche Molecular Biochemicals, Germany) was diluted with HBS buffer to 50ul (120ug / ml) / well; mix A and B, shake gently, and incubate at 15-25°C for 15min ;

[0025] 4. Add the transfection solution drop by drop to each well, shake and mix well; incubate at 37°C for 6 hours;...

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Abstract

The invention relates to the field of microbiological animal cells, in particular to a green fluorescent human liver cancer cell line capable of transfer, wherein high and low transfer mother cells are separated from the high transfer human liver cancer cell lines, realizing pIRES2-EGFP plasmid transfection of the cell strain through liposome transfection method, and high and low transfer cell lines capable of stabilization expression of green fluorescence can be obtained through neomycin extracorporal continuous screening, which are named MHCC97-HG(CGMCC No:063) and MHCC97-LG(CGMCC No:062).

Description

technical field [0001] The invention belongs to the field of microbial animal cell lines, and specifically relates to a human liver cancer cell line capable of spontaneously emitting green fluorescence and capable of transferring, and a method for establishing the same. Background technique [0002] my country is a high-incidence area of ​​liver cancer in the world. About 200,000 people die of liver cancer every year, accounting for about half of the global liver cancer deaths. Modern medicine has conducted scientific research on liver cancer for more than 100 years. Although many achievements have been made, the overall 5-year survival rate of liver cancer patients is still only about 3%. The biggest obstacle to improving the prognosis of liver cancer is its extremely high invasion and metastasis. Even with radical surgical resection, 50% of patients will develop metastasis or recurrence within 5 years. Therefore, in-depth study of the internal mechanism of liver cancer me...

Claims

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Application Information

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IPC IPC(8): A61L27/38C12N5/09C12Q1/00
Inventor 徐泱孙惠川汤钊猷
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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