Method for inducing adult liver stem cells for transforming to high metastatic liver cancer cells and corresponding cells

A technology of liver stem cells and liver cancer cells, applied in tumor/cancer cells, chemical instruments and methods, non-embryonic pluripotent stem cells, etc., can solve the problems of long processing time, poor tumorigenicity, and low rate of spontaneous tumor formation

Active Publication Date: 2017-12-26
SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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Problems solved by technology

It is not yet clear whether malignantly transformed normal cells acquire tumorigenicity and the ability to metastasize
At the same time, the current model for studying lung metastasis of liver cancer is limited. Human-derived liver cancer is poor in lung metastasis, and animal orthotopic liver transplantation is limited to immunodeficient mice, with low success rate and high mortality.
[0006] In the prior art, rat adult liver stem cells that have not been transfected with any gene can spontaneously form tumors after 8-12 cycles of continuous subculture, but the treatment time is long, the spontaneous tumor formation rate is low, and malignant transformation cells without lung metastasizing ability
[0007]

Method used

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  • Method for inducing adult liver stem cells for transforming to high metastatic liver cancer cells and corresponding cells
  • Method for inducing adult liver stem cells for transforming to high metastatic liver cancer cells and corresponding cells
  • Method for inducing adult liver stem cells for transforming to high metastatic liver cancer cells and corresponding cells

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Embodiment 1

[0038] This example provides a method for inducing transformation of adult liver stem cells into highly metastatic liver cancer cells, and provides a stable liver cancer cell with short tumor formation time, good tumorigenicity, and lung metastatic ability.

[0039] The concrete experimental scheme of this embodiment is as follows:

[0040] S1. Expansion and culture of Fisher 344 rat adult liver stem cells WB-F344

[0041](1) Recovery of WB-F344: routinely sterilize the ultra-clean workbench, add 10mL DMEM medium into a sterile 15mL centrifuge tube, take out a tube of frozen rat adult hepatic stem cells WB-F344 from the liquid nitrogen tank, and immediately Place in a water bath at 37°C and oscillate to observe the thawing of the cell suspension. After about 2 / 3 of the thawing, spray the cryopreservation tube with 75% alcohol and put it into an ultra-clean bench. Use a straw to remove the cell suspension. Transfer to a centrifuge tube containing 10mL of medium, blow it evenly...

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Abstract

The invention provides a method for inducing adult liver stem cells for transforming to high metastatic liver cancer cells and the stable liver cancer cells screened by the method. The method comprises the following steps: S1: performing amplification, culture and preservation of mice adult liver stem cells; S2: employing a gene transfection technology for transferring a lentiviral vector containing the Notch1 gene into the mice adult liver stem cells to obtain the mice adult liver stem cells for stable over-expression of the Notch1; S3: using the mice adult liver stem cells for stable over-expression of Notch1, employing a selective condition culture method for culture, performing in-vitro further differentiation, screening the liver cancer cells having strong and stable tumorigenic capability; and S4: employing the screened liver cancer cells, through an in-situ liver transplantation tumor model having same species, liver cancer is formed on liver, double-lung metastasis tumor is spontaneously formed, so that the high metastatic liver cancer cells can be obtained. The screened stable liver cancer cells by the method of the invention have the advantages of short tumor formation time and good tumorigenicity, and have lung metastasis capability.

Description

technical field [0001] The invention relates to a method for inducing transformation of adult liver stem cells into highly metastatic liver cancer cells and corresponding cells. Background technique [0002] Liver cancer is a common malignant tumor in my country. It has the characteristics of insidious onset, rapid progression, easy early hematologic metastasis, difficult to cure, and poor prognosis. Although great progress has been made in the early diagnosis, surgical resection and comprehensive treatment of liver cancer, the five-year survival rate of liver cancer has not improved significantly in the past ten years. Postoperative recurrence of liver cancer has always been the main reason restricting the efficacy of liver cancer treatment. In recent years, it has been found that tumor stem cells are closely related to the formation, development, metastasis, recurrence and drug resistance of tumors. The theory of cancer stem cells believes that most tumors originate from...

Claims

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Application Information

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IPC IPC(8): C12N5/10
CPCC07K14/47C12N5/0693C12N2506/03C12N2510/00
Inventor 刘超施祥德吴文睿许磊波余先焕张锐
Owner SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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