Method and kit for mixed lymphocyte co-culture test detection

A lymphocyte and co-cultivation technology, applied in the field of inspection, to achieve the effect of small sample size, strong popularization and high accuracy

Inactive Publication Date: 2017-09-12
上海众择生物科技有限公司
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to develop a new type of test kit for mixed lymphocyte co-cultivation test f

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
  • Method and kit for mixed lymphocyte co-culture test detection
  • Method and kit for mixed lymphocyte co-culture test detection
  • Method and kit for mixed lymphocyte co-culture test detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The method and kit of the present invention can detect the tissue matching test before organ transplantation of the subject to determine whether the subject is suitable for this organ transplantation and predict the risk of rejection after transplantation. It is usually necessary to obtain fresh peripheral venous blood from organ transplant providers (hereinafter referred to as donors) and organ transplant recipients (hereinafter referred to as recipients) for testing.

[0034] Such as figure 2 As shown, the specific operation steps of this embodiment are as follows:

[0035] The first step, preparation of donor lymphocytes:

[0036] 1) Take fresh peripheral venous blood from the donor and dilute it with PBS at a ratio of 1:1. Slowly trickle along the tube wall and superimpose a test tube filled with 1 / 2 volume of lymphocyte separation solution (be careful not to damage the liquid level stratification), and then centrifuge horizontally at 2000rpm for 20min. The tube...

Embodiment 2

[0052] Whether the subject's T lymphocyte function is low can be detected by the method and kit of the present invention, and it is usually necessary to obtain fresh peripheral venous blood from the subject for testing.

[0053] Such as Figure 4 As shown, the specific operation steps of this embodiment are as follows:

[0054] The first step, the preparation of stimulating cells:

[0055] 1) Commonly used stimulator cells include Epstein-Barr virus-transformed B lymphoblastoid cells (such as N23 cell line, which has been cloned), HTC cells or PBMC cells, etc. Taking N23 cells as an example, take N23 cells in the logarithmic growth phase, centrifuge, resuspend in fresh complete medium, stain with trypan blue, count on a counting plate.

[0056] 2) Add 1 μl of mitomycin (final concentration 10 μg / ml) and 1 ml of PBS to the above cells, bathe in water at 37° C. for 30 minutes, wash with PBS twice by centrifugation.

[0057] 3) Adjust the cell density to 1×10 with RPMI 1640 so...

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 discloses a method and a kit for mixed lymphocyte co-culture test detection. The method and the kit can be used for the mixed lymphocyte co-culture test detection, and are applicable to tissue matching test before organ transplantation, immunologic function test on a low-immunity patient, immunological rejection detection on drugs and transplants, influence detection on T lymphocyte activation caused by other cells or external stimulants, screening of drugs for influencing T cell activation capacity, and other immunoregulation test research. The method and the kit are mainly based on flow cytometry, need a small number of samples, and are easy to operate, high in operability, high in accuracy, high in repeatability, clear in detection result, high in specificity, easy to analyze, low in cost and strong in generalization performance.

Description

technical field [0001] The invention belongs to the technical field of testing, and in particular relates to a method and kit for mixed lymphocyte co-culture test detection. Background technique [0002] When the lymphocytes of two genetically different individuals are mixed and co-cultured in vitro, they will stimulate each other because of the different histocompatibility antigens on their surfaces, which will cause the lymphocytes of the other party to divide, proliferate and transform. This reaction is called two-way mixed lymphocytes. Culture (two way MLC); if one of the lymphocytes is first treated with mitomycine C (mytomycine C) or irradiated to make the DNA in the cells lose their ability to replicate, but still stimulate the transformation of the other lymphocytes, it is called One-way mixed lymphocyte culture (one way MLC). Cell surface antigens encoded by the major histocompatibility complex (MHC) and / or M-locus genes are the main stimuli for this response, and ...

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
IPC IPC(8): G01N33/569C12Q1/02G01N15/14
CPCG01N33/56977C12Q1/02G01N15/10G01N15/14G01N2015/1006G01N2333/52
Inventor 杜飞
Owner 上海众择生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap