Unlock instant, AI-driven research and patent intelligence for your innovation.

DC-CIK (Dendritic cell-Cytokine Induced Killer) cell culture reagent and culture method thereof

A technology of DC-CIK and cell culture, applied in the field of cell culture, can solve the problems of less DC cells, weak antigen sensitization, differentiation, etc., achieve good anti-tumor effect, good cell viability, and promote proliferation

Active Publication Date: 2015-11-18
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
View PDF6 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, there are certain defects in the above-mentioned culture method: during the culture process of DC cells, the initial adherent cells tend to differentiate into macrophages, and few DC cells are harvested; DC cells are sensitized by the serum of tumor patients, and the antigen sensitization is weak, and DC cells The ability of antigen presentation is low, and the killing activity of DC-CIK cells obtained is not high

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • DC-CIK (Dendritic cell-Cytokine Induced Killer) cell culture reagent and culture method thereof
  • DC-CIK (Dendritic cell-Cytokine Induced Killer) cell culture reagent and culture method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1D

[0086] Example 1 DC-CIK cell co-culture example

[0087] (1) Mononuclear cell isolation

[0088]1. Peripheral blood or umbilical cord blood 40mL, centrifuge at 500-800g for 10 minutes, absorb the upper plasma, inactivate at 56°C for 30min, centrifuge at 2000-3000g for 5min, supernatant plasma at 2-8°C for later use;

[0089] 2. For the lower layer of blood, add 2 times the volume of normal saline to resuspend. According to the diluted blood: Ficoll separation solution ratio of 2:1, slowly add the diluted blood to the upper layer of Ficoll separation solution to make the layer clear;

[0090] 3. Centrifuge at 500-700g for 15-30min, and absorb the middle buffy coat layer, which is mononuclear cells.

[0091] (2) DC cell isolation and culture

[0092] 1. Dilute the obtained mononuclear cells with normal saline, centrifuge at 200-400g for 5min, and wash twice;

[0093] 2. Add serum-free medium (containing 5% autologous plasma) to resuspend the cells, press 2-8×10 6 / mL inocula...

Embodiment 2D

[0102] Example 2 DC-CIK cell co-culture example

[0103] (1) Mononuclear cell isolation

[0104] 1. Peripheral blood or umbilical cord blood 40mL, centrifuge at 500-800g for 10 minutes, absorb the upper plasma, inactivate at 56°C for 30min, centrifuge at 2000-3000g for 5min, supernatant plasma at 2-8°C for later use;

[0105] 2. For the lower layer of blood, add 2 times the volume of normal saline to resuspend. According to the diluted blood: Ficoll separation solution ratio of 2:1, slowly add the diluted blood to the upper layer of Ficoll separation solution to make the layer clear;

[0106] 3. Centrifuge at 500-700g for 15-30min, and absorb the middle buffy coat layer, which is mononuclear cells.

[0107] (2) DC cell isolation and culture

[0108] 1. Dilute the obtained mononuclear cells with normal saline, centrifuge at 200-400g for 5min, and wash twice;

[0109] 2. Add serum-free medium (containing 10% autologous plasma) to resuspend the cells, press 2-8×10 6 / mL inocu...

Embodiment 3

[0118] Embodiment 3DC-CIK cell co-culture example

[0119] (1) Mononuclear cell isolation

[0120] 1. Peripheral blood or umbilical cord blood 40mL, centrifuge at 500-800g for 10 minutes, absorb the upper plasma, inactivate at 56°C for 30min, centrifuge at 2000-3000g for 5min, supernatant plasma at 2-8°C for later use;

[0121] 2. For the lower layer of blood, add 2 times the volume of normal saline to resuspend. According to the diluted blood: Ficoll separation solution ratio of 2:1, slowly add the diluted blood to the upper layer of Ficoll separation solution to make the layer clear;

[0122] 3. Centrifuge at 500-700g for 15-30min, and absorb the middle buffy coat layer, which is mononuclear cells.

[0123] (2) DC cell isolation and culture

[0124] 1. Dilute the obtained mononuclear cells with normal saline, centrifuge at 200-400g for 5min, and wash twice;

[0125] 2. Add serum-free medium (containing 15% autologous plasma) to resuspend the cells, press 2-8×10 6 / mL ino...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the technical field of cell culture, and particularly relates to a DC-CIK (Dendritic cell-Cytokine Induced Killer) cell culture reagent and a culture method thereof. The culture reagent comprises DC cell culture reagents, a CIK cell culture reagent and a co-culture reagent, wherein the DC cell culture reagents include a first DC cell culture reagent and a second DC cell culture reagent, the first DC cell culture reagent comprises GM-SCF and IL-4, and the second DC cell culture reagent comprises GM-SCF, IL-4, TNF-alpha and MDC. A large quantity of DC cells and DC-CIK cells can be obtained by adopting the culture method provided by the invention, the proportion of DC-CIK effective cells CD3+CD56+ is high, and the tumor killing capability is strong.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a DC-CIK cell culture reagent and a culture method thereof. Background technique [0002] Tumor is a new organism formed by the body under the action of various tumorigenic factors, and the cells of local tissues lose their normal regulation of their growth at the gene level, resulting in abnormal proliferation and differentiation. Once a new organism is formed, it does not stop growing due to the elimination of the cause. Its growth is not regulated by normal body physiology, but destroys normal tissues and organs. This is especially obvious in malignant tumors. Compared with benign tumors, malignant tumors grow faster and show invasive growth, are prone to bleeding, necrosis, ulcers, etc., and often have distant metastasis, resulting in emaciation, weakness, anemia, loss of appetite, fever, and severe organ damage. Impairment of function, etc., eventually lead to death of...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784C12N5/0783
Inventor 葛啸虎陈海佳王一飞应杰王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD