Culture method of breast cancer MCF-7 cell stem cell levitated spheres

A culture method, MCF-7 technology, applied in the direction of cell culture active agent, tumor/cancer cell, tissue culture, etc., can solve the problem of long formation time of stem cell suspension spheres

Inactive Publication Date: 2017-05-17
FIRST AFFILIATED HOSPITAL OF DALIAN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The technical problem to be solved in the present invention is to overcome the defect that the existing stem cell suspension balls take a long time to form, and provide a method for culturing breast cancer MCF-7 cell stem cell suspension balls

Method used

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  • Culture method of breast cancer MCF-7 cell stem cell levitated spheres
  • Culture method of breast cancer MCF-7 cell stem cell levitated spheres
  • Culture method of breast cancer MCF-7 cell stem cell levitated spheres

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Embodiment

[0025] The human breast cancer MCF-7 cell line in the present invention was purchased from the Shanghai Cell Resource Bank of the Chinese Academy of Medical Sciences

[0026] A culture method of breast cancer MCF-7 cell stem cell suspension ball, is characterized in that, comprises the following steps:

[0027] 1) Collect human breast cancer MCF-7 cells in the logarithmic growth phase, cover 75% to 83% of the culture flask, digest the cells with 0.25% trypsin until the cells become round, and discard the trypsin;

[0028] 2) Add DMEM / H-G complete culture solution to stop the reaction, blow it with a dropper to form a single cell suspension, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, adjust the cell concentration, and sub-culture at 1:3;

[0029] 3) The passaged MCF-7 cells were resuspended in serum-free DMEM / F-12 medium, and 20 ng / mL of EGF, 20 ng / mL of bFGF, 0.4% mass fraction of BSA and 2 % mass fraction of B27, adjust the cell concentration to 0.5 x 10 ...

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Abstract

The invention discloses a culture method of breast cancer MCF-7 cell stem cell levitated spheres. The culture method comprises the following steps: 1) collecting human breast cancer MCF-7 cells at a logarithmic growth phase, paving the collected cells on a 75 to 83 percent culture bottle, dissociating the cells with 0.25 percent by mass pancreatin till the cells become round, and discarding the pancreatin; 2) adding a DMEM / H-G complete culture solution to end a reaction, blowing and beating with a dropper to form a single-cell suspension, centrifuging at the speed of 1,000 revolutions per minute for 5 minutes, discarding supernatant, adjusting the cell concentration, and performing passage in different bottles in the ratio of 1 to 3; 3) re-suspending the MCF-7 cells subjected to passage in a DMEM / F-12 culture medium which does not contain serum, adjusting the cell concentration to 0.5*10<5> / mL, putting into a penicillin glass bottle, covering with a bottle plug, forming an oxygen-lack environment, culturing at the temperature of 37 DEG C in 5% CO2 without substituting the culture medium, mechanically blowing and beating into a single-cell suspension after a microcapsule is formed, and performing conventional passage. The breast cancer MCF-7 cell stem cell levitated spheres are cultured by an oxygen-free culture method, and spherical cell clusters remarkably growing in a suspending way appear on the second day of culture.

Description

technical field [0001] The invention relates to a cell culture method, in particular to a culture method for breast cancer MCF-7 cell stem cell suspension balls. Background technique [0002] There is a small part of tumor stem cells with self-renewal and infinite proliferation ability in the tumor cell population, which is the root cause of tumor occurrence, recurrence, metastasis and failure of conventional treatment. Stem cell suspension balls, commonly known as microspheres, currently have many culture methods, but the formation time is relatively long. Contents of the invention [0003] The technical problem to be solved by the present invention is to overcome the defect that the existing stem cell suspension spheres take a long time to form, and provide a method for culturing breast cancer MCF-7 cell stem cell suspension spheres. [0004] In order to solve the problems of the technologies described above, the present invention provides the following technical soluti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/095C12N5/02
CPCC12N5/0695C12N2501/11C12N2501/115C12N2501/998
Inventor 张斌张霞王艺茜张伟国宋蔚董岩杜福强
Owner FIRST AFFILIATED HOSPITAL OF DALIAN MEDICAL UNIV
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