Bioreactor, stirring paddle thereof and method for cultivating TIL cells through bioreactor

A technology of bioreactor and stirring paddle, applied in the field of cell culture, can solve the problems of unfavorable cost saving and culture space, insufficient clinical treatment, and small scale of TIL cell culture, so as to save medium and culture space and benefit cells Large-scale expansion, which is conducive to the effect of large-scale expansion of cells

Active Publication Date: 2016-05-25
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] The existing TIL cell culture scale is small, the operation is complicated, the steps are cumbersome, the amplification factor is low, the amount obtained by expansion is not enough for clinical treatment, and the culture density should not be too high, which wastes the culture medium, which is not conducive to saving cost and culture space
In addition, the surface antigen expression is low, and the killing effect is poor

Method used

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  • Bioreactor, stirring paddle thereof and method for cultivating TIL cells through bioreactor
  • Bioreactor, stirring paddle thereof and method for cultivating TIL cells through bioreactor

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Embodiment 1

[0029] The stirring paddle was taken out from the 2L bioreactor (purchased from Beijing Yuantang Shengxing Technology Co., Ltd., Bellco, USA), and the medical silicone tube was put on the paddle coat, and after ultraviolet irradiation sterilization, the stirring paddle was reinstalled in the bioreactor. The method of culturing TIL using this bioreactor is as follows:

[0030] Take 2×10 7 PBMC, according to 1×10 6 The density of cells / ml was inoculated in a T75 culture flask with X-VIVO15 medium, added TNF-α solution, IL-2 solution and OKT-3 solution for culture, TNF-α solution, IL-2 solution and OKT-3 solution The amount added is 1% of the medium volume. According to the expansion of the cells, culture in separate flasks. When the total amount of the culture system is greater than 500ml, transfer it to a 2L bioreactor. 2 The ventilation rate is 5%, the dissolved oxygen is 50%, the pH is 7.2, and the stirring speed is 45rpm, and the conditions continue to be cultivated for t...

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Abstract

The invention discloses a bioreactor, a stirring paddle thereof and a method for cultivating TIL cells through the bioreactor. The stirring paddle is wrapped by a material for reducing shear force. The bioreactor comprises the stirring paddle wrapped by the material for reducing shear force, and TIL cells are cultivated through the bioreactor. As TIL cells are cultivated through the bioreactor, and the stirring paddle is wrapped by the material capable of reducing shear force, damage of shear force to cells can be reduced, and mass amplification of cells is facilitated. In the cultivating process, stirring continues, a dynamic system is provided, cells can conduct gas exchanging, the even cultivating environment can be provided for cells, and mass amplification of cells is facilitated. By means of the method, mass amplification of TIL cells can be conducted, the expression quantity of surface antigens of obtained TIL cells is high, and tumor cell killing activity is high.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a bioreactor, a stirring paddle thereof and a method for cultivating TIL cells using the same. Background technique [0002] Tumor infiltrating lymphocytes (tumor infiltrating lymphocytes, TIL cells) are a group of heterogeneous lymphocytes that exist in the tumor stroma. TIL cells have a significant effect on patients with cancerous chest and peritoneal effusions. Generally, lymphocytes isolated from tumor tissue, tumor draining lymph nodes, and cancerous pleural ascites are cultured with IL-2, and their tumor killing activity is MHC-restricted, that is, autologous tumor-specific killer cells with CD3 + CD8 + or CD3 + CD4 + Surface logo. [0003] It has been tested that the anti-tumor effect of TIL cells is 50-100 times that of LAK, and TIL cells can remain in the blood and tumors in the body for up to two months, so it has great potential clinical therapeutic value. At present,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783C12M3/00C12M1/02
CPCC12M27/02C12N5/0636C12N2501/2302C12N2501/25C12N2501/603
Inventor 王一飞陈海佳葛啸虎曾维杰
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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