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Cell culture container capable of improving enrichment efficiency of lung cancer stem cells

A lung cancer stem cell and culture container technology is applied in the field of cell culture containers for improving the enrichment efficiency of lung cancer stem cells.

Pending Publication Date: 2017-05-31
南京九寿堂医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The above method can indeed effectively obtain lung cancer stem cells, but the culture process is cumbersome and the induction efficiency is low. It takes 5-10 days to obtain suspension growth cell spheres, and growth factors need to be added every 2 days before the spheres are formed.

Method used

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  • Cell culture container capable of improving enrichment efficiency of lung cancer stem cells
  • Cell culture container capable of improving enrichment efficiency of lung cancer stem cells
  • Cell culture container capable of improving enrichment efficiency of lung cancer stem cells

Examples

Experimental program
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Effect test

Embodiment 1

[0027] The preparation of embodiment 1 culture bottle of the present invention

[0028] Take a commercially available T-75 culture bottle (domestic Kejing culture bottle, product number 191-203118, air-permeable filter cover, oblique opening, screw opening, sterilized; T-75 means that the available cell culture area is 75cm when laid flat. 2 , this is the common knowledge of those skilled in the art) for subsequent use, on this basis, the laboratory makes the culture bottle provided by the present invention by itself, and the specific steps are as follows:

[0029] Step S1, dissolving γ-polyglutamic acid (sterilized by irradiation with γ-ray cobalt 60, molecular weight 700,000 g / mol) in sterile water to prepare a solution of γ-polyglutamic acid with a concentration of 0.15 mg / mL, and using 0.22 μm membrane filter, spare;

[0030] Step S2, take 15 mL of γ-polyglutamic acid solution and add it into the above-mentioned T-75 culture bottle (the height of the solution is about 0.2...

Embodiment 2

[0035] The preparation of embodiment 2 culture dish of the present invention

[0036] Take a commercially available 60mm cell culture dish (Shanghai Yifei Biotechnology, sterilized; cell growth area 21cm 2 , height is 15mm, this is the common knowledge of those skilled in the art) for subsequent use, on this basis, the cell culture dish provided by the present invention is self-made in the laboratory, and the concrete steps are as follows:

[0037] Step S1, dissolving gamma polyglutamic acid (sterilized by gamma ray cobalt 60) in sterile water to prepare a gamma polyglutamic acid solution with a concentration of 0.15 mg / mL, filtering it with a 0.22 μm filter membrane, and set aside;

[0038] Step S2, take 5mL of γ-polyglutamic acid solution and add it to the above 60mm cell culture dish (the solution height is about 0.24cm), shake well to make it evenly cover the bottom of the entire cell culture dish, put it in a 37°C incubator and let it stand 5 hours;

[0039] Step S3, ta...

Embodiment 3

[0043] Embodiment 3 The preparation of culture plate of the present invention

[0044] Take a commercially available 6-well plate (domestic Membrane, sterilized; single hole area 9.5cm 2 , single hole volume 16.5mL, this is the common knowledge of those skilled in the art) standby, on this basis, the cell culture dish provided by the present invention is self-made in the laboratory, and the specific steps are as follows:

[0045]Step S1, dissolving gamma polyglutamic acid (sterilized by gamma ray cobalt 60) in sterile water to prepare a gamma polyglutamic acid solution with a concentration of 0.15 mg / mL, filtering it with a 0.22 μm filter membrane, and set aside;

[0046] Step S2, take 2.5mL of γ-polyglutamic acid solution and add it to the above 6-well plate (the height of the solution is about 0.26cm), shake well to make it evenly cover the bottom of the entire cell culture well, put it in a 37°C incubator and let it stand 5 hours;

[0047] Step S3, take out the culture pl...

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Abstract

The invention discloses a cell culture container capable of improving the enrichment efficiency of lung cancer stem cells. The cell culture container comprises a culture container main body, and an enveloping layer is arranged on the inner wall of the culture container main body and is made of gama-polyglutamic acid. The culture container with the gama-polyglutamic acid containing enveloping layer can obviously improve the formation efficiency of suspended grown cell spheres, ideal suspended growing cell spheres can be formed within 24 hours, the lung cancer stem cells can be highly enriched in the suspended growing cell spheres, and the cell culture container has higher efficiency and more convenience compared with the existing serum culture medium-free cell growth factor method.

Description

technical field [0001] The invention belongs to the field of stem cells, and in particular relates to a cell culture container for improving the enrichment efficiency of lung cancer stem cells. Background technique [0002] Lung cancer is one of the most harmful malignant tumors to human health and life. So far, the existing lung cancer treatment methods can only prolong the survival period of patients and improve their quality of life, and almost all patients still die from the primary disease in the end. In recent years, some scholars have proposed that there is a very small number of stem cell populations with unlimited proliferation potential and self-renewal ability in tumor tissues, which play a vital role in the occurrence and development of tumors. Targeted therapy for this part of cells will have a profound impact on cancer treatment. However, the content of cancer stem cells (CSCs) in tumor tissues is extremely small, which has become a major obstacle restricting...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12M1/24C12M1/22
CPCC12M23/08C12M23/10
Inventor 郭守河张康
Owner 南京九寿堂医药科技有限公司
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