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In-vitro activation method of cytokine-induced killer cells

A technology for killing cells and cytokines, which is applied in the field of biomedicine and can solve the problems of not increasing the proportion of NK cells and lack of functional cells.

Active Publication Date: 2020-10-30
北京科霖恩生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these changes did not increase the proportion of NK cells, and a functional cell was missing in the culture system

Method used

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  • In-vitro activation method of cytokine-induced killer cells
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  • In-vitro activation method of cytokine-induced killer cells

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Embodiment 1

[0040] This embodiment discloses a method for in vitro activation and culture expansion of cytokine-induced killer cells (CIK cells). Taking 40ml of human peripheral blood as an example, the method is as follows:

[0041] 1. Cell Culture and Harvesting

[0042] (1) Preparation of 2% gelatin: Weigh 2 g of gelatin (product of Sigma, USA, product number G7041), weigh 0.85 g of sodium chloride, and add 90 ml of deionized water. Stir to dissolve, add deionized water to adjust the total volume to 100ml. Filter through a 0.22 μm syringe filter. Store at 4°C.

[0043] (2) Coating of activator: prepare two bottom areas of 75cm 2 The culture flasks are marked as ctr and test respectively. Add physiological saline to the culture bottle marked as ctr, add 10ml of 2% gelatin (activator) to the culture bottle marked as test, and spread it on the bottom of the culture bottle. Place the flask at 37°C for 2 hours.

[0044] It should be noted that ctr represents a traditional method, and ...

Embodiment 2

[0081] This embodiment discloses a method for in vitro culture and expansion of cytokine-induced killer cells (CIK cells). Taking 50ml of human peripheral blood as an example, the method is as follows:

[0082] 1. Cell Culture and Harvesting

[0083] (1) Coating with activator: prepare 4 pieces with a bottom area of ​​75cm 2 The culture flasks are marked as ctr and test respectively. Add physiological saline to the culture bottle marked as ctr, add 10ml of 2% gelatin (activator) to the culture bottle marked as test, and spread it on the bottom of the culture bottle. Place the flask at 4°C overnight.

[0084] (2) Activator treatment: suck off the gelatin in the culture bottle marked as test, add physiological saline, shake gently to spread the salt level on the bottom of the culture bottle, and suck out the normal saline. Aspirate the normal saline in the culture flask marked as ctr. The culture bottle is ready for use.

[0085] (3) Take 50ml of peripheral blood from healt...

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Abstract

The invention discloses an in-vitro activation method of cytokine-induced killer cells, and relates to the field of biological medicines. Specifically, according to the method, before cell culture, aculture bottle is coated with a gelatin solution with the concentration of 0.1-2%, then the gelatin solution is sucked away, and cells are inoculated. Compared with an existing patent method, gelatincoating is conducted before CIK cells are cultured, the ratio of target cells to natural killer cells can be remarkably increased, and the killing activity of the cells to tumor cells is enhanced. Themethod is not only suitable for CIK cell culture, but also suitable for natural killer cell culture, and has great market prospect and economic value.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an in vitro activation method of cytokine-induced killer cells. Background technique [0002] Cancer is a disease that seriously threatens human health. As people live longer, cancer has become a common and frequently-occurring disease, and it is also a difficult problem that clinicians often face. Cancer treatments include surgery, chemotherapy, radiation therapy, and biological therapy. As a relatively novel means of cancer treatment, biological therapy is gradually entering clinical application from the clinical trial stage, and has become an indispensable way of cancer treatment. Cytokine-induced killer cells (CIK cells) are also a new clinical approach. In Western Europe, CIK cell therapy has been approved by the government for the treatment of acute leukemia and myelodysplastic syndrome. my country has also carried out a number of clinical trials for the treatment of...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0638C12N5/0646C12N2533/54C12N2501/24C12N2501/2302C12N2501/2315C12N2501/515
Inventor 郭子宽潘慧欣王恒湘哈小琴
Owner 北京科霖恩生物科技有限公司
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