Method for in vitro expanding tumor stem cells

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...

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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 determined in the previous step, and finally after embedding In vitro three-dimensional culture of tumor cells to achieve in vitro expansion of tumor stem cells, but due to the differentiation of tumor stem cells during the culture process, other cells with less stemness are formed, and cells differentiated from tumor stem cells can grow in the culture medium Normal growth in the tumor cells, so the final cell population will include some differentiated cells, and make the quality of tumor stem cells poor

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