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Preparation method for enhanced CIK (cytokines-induced killer) cells and application of rhizoma atractylodis macrocephaiae polysaccharide and lycium barbarum polysaccharide

A technique for Atractylodes Rhizoma polysaccharide and Lycium barbarum polysaccharide, which is applied in the field of biomedicine, can solve problems such as limitations, and achieve the effects of improving tumoricidal activity, improving tumoricidal effect, and improving proliferation activity.

Pending Publication Date: 2017-09-01
南华生物医药股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] CIK cells play an important role in immune regulation and anti-tumor immunotherapy, but their therapeutic effect is closely related to cell infusion dose, effector cell ratio and cytotoxicity to tumor cells, while the proliferation and tumoricidal activity of CIK cells depend on a variety of The stimulation of cytokines and exogenous factors, so the clinical application of CIK cells is limited by their in vitro proliferation activity and in vivo tumoricidal activity, and how to improve their proliferation efficiency and toxicity in vitro is the primary problem to be solved

Method used

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  • Preparation method for enhanced CIK (cytokines-induced killer) cells and application of rhizoma atractylodis macrocephaiae polysaccharide and lycium barbarum polysaccharide
  • Preparation method for enhanced CIK (cytokines-induced killer) cells and application of rhizoma atractylodis macrocephaiae polysaccharide and lycium barbarum polysaccharide
  • Preparation method for enhanced CIK (cytokines-induced killer) cells and application of rhizoma atractylodis macrocephaiae polysaccharide and lycium barbarum polysaccharide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1. Density gradient centrifugation to separate human peripheral blood mononuclear cells (PBMCs), the steps are:

[0034] 1) Peripheral blood collection: Professional medical blood collection personnel are invited to collect 30 mL of blood from healthy volunteers through venous blood collection into vacuum blood collection tubes containing heparin sodium anticoagulant, and repeatedly invert the blood collection tubes several times.

[0035] 2) Collect serum: transfer the blood to a 50 mL centrifuge tube, centrifuge at 1000g for 10 min, and form two phases of upper serum and lower layer of cells after centrifugation is completed. Collect the upper layer of plasma into a new centrifuge tube, mark it, inactivate it in a 56-degree water bath for 30 min, and set it aside.

[0036] 3) Sample dilution: Add normal saline to dilute the remaining cell layer to a volume of 30 mL, and pipette to evenly pipette.

[0037] 4) Gradient centrifugation: Take another 50 mL centrifuge tube...

Embodiment 2

[0052] 1. Density gradient centrifugation to separate human peripheral blood mononuclear cells (PBMCs), the steps are:

[0053] 1) Peripheral blood collection: Professional medical blood collection personnel are invited to collect 30 mL of blood from healthy volunteers through venous blood collection into vacuum blood collection tubes containing heparin sodium anticoagulant, and repeatedly invert the blood collection tubes several times.

[0054] 2) Collect serum: transfer the blood to a 50 mL centrifuge tube, centrifuge at 1000g for 10 min, and form two phases of upper serum and lower layer of cells after centrifugation is completed. Collect the upper layer of plasma into a new centrifuge tube, mark it, inactivate it in a 56-degree water bath for 30 min, and set it aside.

[0055] 3) Sample dilution: Add normal saline to dilute the remaining cell layer to a volume of 30 mL, and pipette to evenly pipette.

[0056] 4) Gradient centrifugation: Take another 50 mL centrifuge tube...

Embodiment 3

[0074] 1. Density gradient centrifugation to separate human peripheral blood mononuclear cells (PBMCs), the steps are:

[0075] 1) Peripheral blood collection: Professional medical blood collection personnel are invited to collect 30 mL of blood from healthy volunteers through venous blood collection into vacuum blood collection tubes containing heparin sodium anticoagulant, and repeatedly invert the blood collection tubes several times.

[0076] 2) Collect serum: transfer the blood to a 50 mL centrifuge tube, centrifuge at 1000g for 10 min, and form two phases of upper serum and lower layer of cells after centrifugation is completed. Collect the upper layer of plasma into a new centrifuge tube, mark it, inactivate it in a 56-degree water bath for 30 min, and set it aside.

[0077] 3) Sample dilution: Add normal saline to dilute the remaining cell layer to a volume of 30 mL, and pipette to evenly pipette.

[0078] 4) Gradient centrifugation: Take another 50 mL centrifuge tube...

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Abstract

The invention relates to the field of biological medicines, in particular to a method for enhancing activity of CIK (cytokines-induced killer) cells through traditional Chinese medicine induction. The method comprises the step of adding rhizoma atractylodis macrocephaiae and lycium barbarum extracts into a culture fluid for the CIK cells, wherein the rhizoma atractylodis macrocephaiae and lycium barbarum extracts are rhizoma atractylodis macrocephaiae polysaccharide and lycium barbarum polysaccharide. According to the method, the activity of the CIK cells in the aspects of proliferating, tumor killing and immunity can be activated; the in-vitro tumour killing effect of the CIK cells is improved, and meanwhile, a large number of CIK cells can be obtained, so that the cell culture time is shortened, and the immunological activity of the CIK cells can also be enhanced; the enhanced CIK cells prepared by utilizing the method can be used for treating tumour diseases.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for preparing enhanced CIK cells induced by a combination of traditional Chinese medicine extracts and its application in CIK functional activity. Background technique [0002] Malignant tumors seriously endanger human health. Conventional surgery, radiotherapy, chemotherapy and other treatment methods cannot completely eliminate cancer cells in the body. Adoptive immunotherapy, which reinfuses immune active cells into the body, can directly kill tumor cells, and can also regulate and enhance the body's immune system. Therefore, it has become an important auxiliary means of tumor treatment. [0003] Immune cell therapy includes NK, LAK, CIK, DC-CIK, TIL, etc. Among them, cytokine-induced killer cells (CIK cells) are a very promising adoptive immune cell discovered in recent years. It has both the strong anti-tumor activity of T lymphocytes and the non-MH...

Claims

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Application Information

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IPC IPC(8): C12N5/0783A61K35/17A61P35/00
CPCA61K35/17C12N5/0638C12N2501/515C12N2501/2302C12N2500/76C12N2500/34
Inventor 向双林黄招琴颜峰彭海宁杨满君王怡雅刘京韬胡翔
Owner 南华生物医药股份有限公司
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