Efficient CIK cell culturing method

A cell culture and high-efficiency technology, which is applied in animal cells, vertebrate cells, blood/immune system cells, etc., can solve the problems of low efficiency, improve the culture speed, increase the highest apoptosis rate, increase the number and speed of amplification Effect

Active Publication Date: 2015-06-03
中科聚研干细胞有限公司
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Problems solved by technology

[0005] The efficiency of the CIK culture method adopted by many researchers and institutions is low. The number of peripheral blood mononuclear cells extracted from 70-100ml of fresh blood only reached 10 after about 14 days of induction culture. 9 Magnitude

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  • Efficient CIK cell culturing method
  • Efficient CIK cell culturing method
  • Efficient CIK cell culturing method

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Embodiment Construction

[0023] The present invention will be further described in conjunction with the comparative experiment between adopting conventional CIK cell culture method and the efficient CIK cell culture method used in the present invention below:

[0024] A high-efficiency CIK cell culture method is characterized in that the culture steps are:

[0025] Step 1: Monocyte Isolation

[0026] Put the quantitative blood sample into a centrifuge tube, centrifuge the blood sample, and discard the rest at the bottom; dilute the obtained blood cells with normal saline. After centrifugation, the supernatant was discarded; the buffy coat cells were slowly sucked into a new centrifuge tube, and physiological saline was added to the centrifuge tube, and mixed evenly to obtain a white blood cell dilution, and finally obtained The white blood cell dilution is 70%-80% of the blood sample volume, centrifuge the white blood cell dilution, and discard the supernatant; resuspend with 5%-10% of the blood samp...

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Abstract

The invention relates to the field of cytobiology and particularly discloses an efficient CIK cell culturing method by means of efficiently culturing the CIK cells through the steps of monocyte separation, culture bottle inoculation, culture bag inoculation and cell collection, adding the inducible factors such as IL-1, IL-2, IL-24, IFN-gamma, CD3AK, EGF during the culture process, inducing the peripheral blood mononuclear cell into the CIK cell and performing in vitro efficient amplification. After culturing for 10 days, the peripheral blood mononuclear cell with the initial cell number of 2*10<6> is induced and amplified to the 1*10<10> CIK cell, the CIK cell in vitro multiplication culture speed and the final obtained cell number can be effectively increased, besides, by adopting the CIK cell cultured by the method, the maximum apoptosis rates of the cervical cancer Hela cell and the myeloma Sp20 cell are 64.2% and 54.3%.

Description

technical field [0001] The invention belongs to the field of cell biology, and specifically relates to a cell that can effectively increase the speed of in vitro expansion and culture of CIK cells and finally obtain them by adding IL-1, IL-2, IFN-γ, CD3 AK, and EGF as inducing factors. The number and the number of CIK cells cultivated can greatly increase the highest apoptosis rate of cervical cancer Hela cells and myeloma Sp20 cells. A high-efficiency CIK cell culture method. Background technique [0002] With the in-depth research and development of cell biology technology and clinical medical technology, the rapid development of cell culture technology has been further promoted. However, the research on the cultivation of autologous immune cells is limited, which limits the technical development and application of immune cells to treat various diseases. [0003] Cytokine-Induced Killer (CIK) is a new type of immunocompetent cell. CIK cells have strong proliferation abilit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
Inventor 刘慧玉李春雨李鹏高妍刘法显
Owner 中科聚研干细胞有限公司
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