Combined formula kit for analyzing phenotype and function of CD1c+ dendritic cell subsets and application thereof

A dendritic cell and kit technology, which is applied in the biological field, can solve the problem of not being able to study CD1c+DC subgroups, etc., and achieve the effect of rapid one-step determination, long time consumption, and saving a lot of time.

Active Publication Date: 2019-10-29
BLUE ELEGANT BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the above-mentioned CD11c+CD11b+DC subgroup detection kit cannot meet the needs of researching CD1c+DC subgroup

Method used

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  • Combined formula kit for analyzing phenotype and function of CD1c+ dendritic cell subsets and application thereof
  • Combined formula kit for analyzing phenotype and function of CD1c+ dendritic cell subsets and application thereof
  • Combined formula kit for analyzing phenotype and function of CD1c+ dendritic cell subsets and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Pretreatment of Non-small Cell Lung Cancer Patient / Healthy People's Peripheral Blood

[0055] The preprocessing steps are as follows:

[0056] (1) Take 1 drop (10-100 μl) of venous peripheral blood from patients with non-small cell lung cancer and healthy adults, and perform anticoagulant treatment;

[0057] (2) Mix peripheral whole blood with 2 ml of 1× red blood cell lysate (Biolegend), rotate and oscillate for 10 seconds, and let stand at room temperature in the dark for 15 minutes;

[0058] (3) centrifuge (350g * 5 minutes) with centrifuge, pour over the supernatant, then the precipitated cells are suspended in 2 milliliters of cell staining fluid (containing the PBS solution of 2.5% fetal bovine serum);

[0059] (4) Leukocyte-stimulating factor (BD) was added at a concentration ratio of 0.1%, and the cells were incubated at a constant temperature of 37 degrees for 6 hours for later use.

Embodiment 2

[0060] Example 2 Analysis of the degree of development and differentiation of CD1c+ dendritic cell subsets in peripheral blood of non-small cell lung cancer patients / healthy people

[0061] The spare blood cells were centrifuged (350 g) for 5 minutes, the supernatant was decanted, and the cells were suspended in 100 μl of cell staining solution. Then add 2 μl of anti-human CD1c and CD40 antibodies (Biolegend), incubate at room temperature for 30 minutes, then add 2 ml of cell staining solution, and repeat centrifugation twice at 350 g for 5 minutes each time. After decanting the supernatant, fix the cells with 2 ml of 2% formalin solution, and incubate at room temperature in the dark for 20 minutes for later use.

Embodiment 3

[0062] Example 3 Functional Analysis of Peripheral Blood CD1c+ Dendritic Cell Subsets of Non-small Cell Lung Cancer Patients / Healthy People

[0063] (1) Suspend the fixed spare cells in 2 ml of cell penetration solution (Biolegend), then repeat centrifugation (350g) for 10 minutes twice;

[0064] (2) Resuspend the pelleted cells after centrifugation in 100 μl of cell permeation buffer, then add 2 μl of IL-6 (Biolegend) and IL-10 antibody (BD), and incubate at room temperature for 30 minutes in the dark;

[0065] (3) The incubated cells were suspended in 2 ml of cell penetration solution, and then centrifuged (350g) for 5 minutes twice;

[0066] (4) Finally, decant the supernatant, resuspend the pelleted cells in 0.5 ml of cell staining solution, and analyze and test with a flow cytometer.

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Abstract

The invention provides a combined formula kit for analyzing the phenotype and function of CD1c+ dendritic cell subsets and an application thereof. The test subjects of the combined formula kit for analyzing the phenotype and function of the CD1c+ dendritic cell subsets include CD1c, CD40, IL-6 and IL-10. The combined formula kit for analyzing the phenotype and function of the CD1c+ dendritic cellsubsets provided by the invention can be used for efficiently and rapidly identifying the phenotype of the CD1c+ dendritic cell subsets in peripheral blood and simultaneously analyzing the function; the accuracy rate is guaranteed, and the economic cost due to the detection of large amounts of surface antigen molecules is reduced; and the test method is simple and easy.

Description

technical field [0001] The invention belongs to the field of biotechnology, and uses flow cytometry to perform immunodetection and analysis of human peripheral blood dendritic cells, in particular to a combined formula kit for analyzing the phenotype and function of CD1c+ dendritic cell subgroups and its application. Background technique [0002] Flow cytometry analysis technology has been used in clinical and scientific research as the main technology of immunology. Dendritic cells, as the main regulatory cells of the human immune system, are one of the hotspots in immunology research. At present, the detection of dendritic cells mainly relies on flow cytometry analysis technology, but the current routine cytometry analysis schemes for the determination of dendritic cells are various, and there is no unified and standardized mode. This is mainly because the research on dendritic cells is changing with each passing day, and many different subtypes of dendritic cells have b...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N33/574
CPCG01N33/533G01N33/57496G01N33/56972G01N2333/70578G01N2333/5412G01N2333/5428
Inventor 周放陈小平秦莉古艳丽许文龙卢永常旭韦国建容志恩
Owner BLUE ELEGANT BIOTECH CO LTD
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