Liver-cancer stem cell, its separation and use

A liver cancer stem cell and separation method technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve problems such as metastasis, tumor recurrence, tumor mass shrinkage, etc.

Inactive Publication Date: 2007-04-04
SHANGHAI INST OF ONCOLOGY
View PDF0 Cites 14 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, due to the concept of "cancer stem cell (CSC)" [Reya et al., Nature (2001) 414: 105-111], people's concept of evaluating the effect of tumor treatment has changed significantly. Good conventional chemotherapy drugs can only kill the "common tumor cells" that account for a large proportion of the tumor tissue, and the tumor mass can be significantly reduced or even "disappeared" during treatment. One) of the "very tumor cells" - cancer stem cells are helpless [Bissell et al, Cancer Cell (2005) 7: 17-23; Patrawala et al, Cancer Research (2005) 65: 6207-6219], and there are signs that After conventional chemotherapy drugs treat tumors, because the balance between cancer stem cells and normal tumor cells is broken, it will stimulate cancer stem cells to divide asymmetrically or differentiate into normal tumor cells, eventually leading to tumor recurrence or even metastasis

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Liver-cancer stem cell, its separation and use
  • Liver-cancer stem cell, its separation and use
  • Liver-cancer stem cell, its separation and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Isolation and identification of cancer stem cells in liver cancer cell lines

[0059] The specific AC133 / CD133 / 1 monoclonal antibody and magnetic bead sorting system enrich and separate stem cells from liver cancer cell lines:

[0060] (1) Expression of stem cell marker CD133 in liver cancer cell lines: Figures 1a-f are human liver cancer cell lines, including (a) SMMC7721, (b) BEL7402, (c) Huh-7, (d) Hep3B, (e) MHCC-LM3, (f) MHCC-97L, immunocytochemical test results show that there are a small number of CD133+ cells in the above cell lines (arrow);

[0061] (2) CD133 efficiently enriched by magnetic bead sorting + Cells, of which (g) pre-sorting cells CD133 + The proportion of positive cells is low, (h) CD133 after sorting + The positive rate is greatly increased .

[0062] Western blot (i) results also confirmed this point, where "+" means CD133 after sorting + Are positive cells, "-" means CD133 after sorting - It is a negative cell, and β-actin is an internal control.

Embodiment 2

[0064] Identification and identification of cancer stem cells in human liver cancer-related tissues

[0065] The expression of stem cell marker CD133 in paraffin-embedded liver cancer, adjacent liver, liver cirrhosis, and normal liver tissue sections (immunohistochemical detection): Figure 2a-c is liver cancer tissue, di is adjacent liver tissue, jl is liver cirrhosis tissue , M, n are normal liver tissues.

[0066] It can be seen from Figure 2 that in addition to normal liver tissues, there are few CD133-positive cells (yellow-brown) in liver cancer, para-cancerous liver, and cirrhosis tissues, and the white arrow indicates the Hering’s canal-like structure.

Embodiment 3

[0068] CD133 + In vitro cloning ability detection of liver cancer stem cells

[0069] CD133 was detected by soft agar colony formation assay + The cloning ability of liver cancer stem cells in soft agar, the specific steps are as follows:

[0070] (1) Pave the 6-well cell culture plate (bottom layer) with 0.6% low melting agar-stem cell culture medium containing 10% fetal bovine serum prepared at 42°C;

[0071] (2) After curing at 37℃, use 5×10 3 CD133 + Or CD133 - SMMC7721-GFP single cell-soft agar suspension covers the bottom glue;

[0072] (3) Solidify at 37°C, add 2ml of stem cell culture medium to each well, and change the fresh medium every 5 days;

[0073] (4) Three weeks later, clones with more than 15 cells were counted and photographed under an inverted fluorescence microscope, and the data was statistically analyzed.

[0074] The results are shown in Figure 3: The bar graph (a) shows the number of colonies formed, expressed as the average number ± sd (n = 12 low power f...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

A liver cancer stem cell, its separation and use are disclosed. The process is carried out by sorting by specific AC133 / CD133 / 1 single-clone antibody and magnetic ball, collecting liver-cancer stem cell and separating to obtain the CD133+ liver cancer stem cell. It can be used to screen effective anti-cancer medicine or medicinal target, diagnose and treat bi-functional specific single-clone antibody of liver cancer.

Description

Technical field [0001] The present invention relates to stem cells, in particular to a liver cancer stem cell. Background technique [0002] With the advancement of science and technology, great progress has been made in the research of tumor diagnosis and treatment. In particular, the introduction of advanced modern diagnostic equipment such as CT, NMR, and PET has made the diagnosis of tumors "accurate" and "early" advanced. A big step. Also in terms of treatment, due to the introduction of various new biological treatments and chemotherapeutics in recent years, the drug treatment effect of tumors has also been significantly improved, but so far the occurrence mechanism of many tumors (including liver cancer) has not been clarified. In recent years, due to the concept of "cancer stem cell (CSC)" [Reya et al., Nature (2001) 414:105-111], people’s concept of evaluating the effect of tumor treatment has changed significantly. Good conventional chemotherapeutics can only kill the "...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N5/08A61K35/12A61P35/00C07K16/18C12Q1/04A61K39/395C12N5/095
Inventor 李锦军葛超顾健人
Owner SHANGHAI INST OF ONCOLOGY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap