Method for in vitro expanding tumor stem cells

A tumor stem cell, in vitro expansion technology, applied in the direction of tumor/cancer cells, animal cells, cell culture medium, etc., can solve the problem of poor quality of tumor stem cells, and achieve the effect of increasing quantity and quality

Inactive Publication Date: 2017-05-10
浙江译美生物科技有限公司
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0003] At present, the Chinese patent with the publication number CN103571792A discloses a method for expanding tumor stem cells in vitro and a processing method thereof. It first determines the matrix stiffness of the environment in which the tumor stem cells are expanded in vitro, and then uses alginate to Gel scaffold embedding requires in vitro expansion of tumor cells, so that the matrix stiffness of the embedded gel scaffold is 90-110% of the matrix stiffness of the tumor stem cell environment det

Method used

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  • Method for in vitro expanding tumor stem cells

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Embodiment 1

[0022] The processing method of the method for expanding tumor stem cells in vitro in Example 1

[0023] Step 1: Take ordinary cell culture medium, add lung cancer tissue cells to the culture medium, and culture under sterile conditions at 37° C. for 30 h.

Embodiment 2

[0024] The processing method of the method for expanding tumor stem cells in vitro in Example 2

[0025] Step 1: Take serum-free medium, and add lung cancer tissue cells to the medium;

[0026] Step 2: Add 20 mg / L IFO to the culture medium, and incubate at 37°C for 25 hours under sterile conditions.

Embodiment 3

[0027] The processing method of the method for expanding tumor stem cells in vitro of embodiment 3

[0028] Step 1: Take serum-free medium, and add 10mg / L IFO and 2mg / L MPF to the medium;

[0029] Step 2: Put the lung cancer tissue cells into the medium, add 0.5 mg / L Accutase at the same time, and stir, then add it every four hours, and change the direction of stirring;

[0030] Step 3: Put the culture medium under sterile conditions at 37° C. for 28 hours.

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Abstract

The invention discloses a method for in vitro expanding tumor stem cells. The invention aims to solve the problem that the stem cell quality is poor according to a traditional method for in vitro expanding tumor stem cells. According to the technical scheme, the method for in vitro expanding tumor stem cells comprises the following steps: 1) taking a serum-free medium and adding 20-35mg/L B27, 10-25mg/L IFO and 1-3mg/L MPF to the serum-free medium; 2) increasing the temperature of the medium to 37 DEG C and sterilizing with ultraviolet rays; 3) transferring the tumor tissues into the medium under an aseptic condition and stirring along a same direction; 4) adding 0.5mg/L Accutase into the medium acquired in the step 3) every 4 hours, wherein 0.1mg/L Accutase is additionally added at each time and the stirring direction is reversely adjusted when the Accutase is added each time; 5) culturing for 20-35h and then centrifugally separating, thus obtaining the tumor stem cells. The quality of stem cells obtained according to the method for in vitro expanding tumor stem cells is higher.

Description

technical field [0001] The present invention relates to the field of cell culture, and more specifically, relates to a method for expanding tumor stem cells in vitro. Background technique [0002] Cancer stem cells are cells with the ability of "Self-Renewal" and "Differentiation". Generally, these cells are considered to have the potential to form tumors and develop into cancer, especially as cancer After metastases, the source of new types of cancer is generated. Since the number of tumor stem cells is very small, it is difficult to be found, and it is also difficult for anticancer drugs to have an effect on it. Therefore, in order to conquer tumors as soon as possible, it is necessary to conduct research on tumor stem cells. The technology of expanding tumor stem cells in vitro has emerged as the times require. [0003] At present, the Chinese patent with the publication number CN103571792A discloses a method for expanding tumor stem cells in vitro and a processing metho...

Claims

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Application Information

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IPC IPC(8): C12N5/095
CPCC12N5/0695C12N5/0037C12N2500/82C12N2500/90C12N2501/06C12N2501/998
Inventor 陈继冰吴振化
Owner 浙江译美生物科技有限公司
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