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Induced proliferation method of immune cell

A technology of immune cells and cells, applied in the field of induction and expansion of immune cells, can solve the problems of limited application of cell immunotherapy, poor uniformity of immune cells, high probability of tumorigenesis, etc., and achieve the improvement of induction differentiation efficiency and T cell expansion Efficiency improvement and uniformity improvement effect

Active Publication Date: 2018-06-22
广州沙艾生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, in the prior art, human autoimmune cells obtained through in vitro culture have the problems of low expansion efficiency, poor uniformity of immune cells, reinfusion into patients, and high probability of tumorigenesis.
Applications of cellular immunotherapy therapies are limited

Method used

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  • Induced proliferation method of immune cell
  • Induced proliferation method of immune cell
  • Induced proliferation method of immune cell

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0027] In this example, the inventors will describe in detail the method for inducing and expanding immune cells, including inducing and expanding isolated mononuclear cells into immune cells, and detecting the activity of immune cells.

[0028] 1. Experimental method

[0029] 1. Preparation of Anti-human CD16 Coated ZellWerk Bioreactors

[0030] 1.1 Add 15 mL of 2.5 μg / mL anti-human CD16 monoclonal antibody dissolved in medical normal saline to the bioreactor, shake the culture bottle gently to make the antibody cover the culture surface, and overnight at 4°C in the dark.

[0031] 1.2 Recover the antibody coating solution before use, wash the bioreactor once with 15mL of normal saline, and then use 15mL of T cell expansion medium (OpTmizer TM CTS TM T-cell expansion SFM) and washed once.

[0032] 2. Collect peripheral blood, separate peripheral blood plasma and mononuclear cells

[0033] 2.1 Collect about 100ml of human peripheral blood with a sterile blood collection bag...

Embodiment 2

[0076] In this example, the inventors will describe in detail the method for inducing and expanding immune cells, including inducing and expanding isolated mononuclear cells into immune cells, and detecting the activity of immune cells.

[0077] 1. Experimental method

[0078] 1. Preparation of Anti-human CD16 Coated ZellWerk Bioreactors

[0079] 1.1 Add 15 mL of 2.5 μg / mL anti-human CD16 monoclonal antibody dissolved in medical normal saline to the bioreactor, shake the culture bottle gently to make the antibody cover the culture surface, and overnight at 4°C in the dark.

[0080] 1.2 Recover the antibody coating solution before use, wash the bioreactor once with 15mL of normal saline, and then use 15mL of T cell expansion medium (OpTmizer TM CTS TM T-cell expansion SFM) and washed once.

[0081] 2. Collect peripheral blood, separate peripheral blood plasma and mononuclear cells

[0082] 2.1 Collect about 100ml of human peripheral blood with a sterile blood collection b...

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Abstract

The invention provides an induced proliferation method of an immune cell. The method comprises the steps of: (1) inoculating a mononuclear cell into a bioreactor for cultivate for a day under an aseptic condition in 2-7% of CO2 and 8-10% of O2 at 37 DEG C, (2) continuously culturing the mononuclear cell cultured in step (1) for 14-21 days to form the immune cell, wherein an anti-human CD61 antibody is coated in advance in the bioreactor; the bioreactor comprises a first culture medium; the first culture medium is a T cell proliferation culture medium comprising 10% autologous plasma, 1000IU / mlIL-2 and 0.01KE / ml sapylin; the continuous culture from day 1 to day 8 in step (2) is performed in a second culture medium at 37 DEG C under the aseptic condition in 2-7% of CO2 and 8-10% of O2; thesecond culture medium is a T cell proliferation culture medium comprising 10% autologous plasma and 1000IU / ml IL-2, or a T cell proliferation culture medium comprising 1000IU / ml IL-2; the continuous culture from day 9 to day 21 is performed in a third culture medium at 37 DEG C under the aseptic condition in 12-18% of O2 and 2-7% of CO2; and the third culture medium is a SuperCulture TM L500 culture medium comprising 1000IU / ml IL-2.

Description

technical field [0001] The invention relates to the field of biology, in particular, the invention relates to a method for inducing and expanding immune cells. Background technique [0002] Cellular immunotherapy is to collect the body's own immune cells, culture them in vitro, increase their number by thousands of times, and enhance their targeted killing function, and then infuse them back into the human body to kill pathogens, cancer cells, The mutated cells break immune tolerance, activate and enhance the body's immune ability, and take into account the dual effects of treatment and health care. [0003] However, in the prior art, there are problems such as low expansion efficiency and poor uniformity of immune cells obtained by culturing the obtained human immune cells in vitro, and high probability of tumorigenesis when reinfused into patients. The application of cellular immunotherapy therapy is limited. [0004] Therefore, how to further effectively improve the exp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0786C12N5/0783
CPCC12N5/0636C12N5/0645C12N2500/30C12N2501/2302C12N2501/599C12N2509/00
Inventor 徐智峰张新
Owner 广州沙艾生物科技有限公司
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