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SN/T 2497.24-2010: Test method of import and export dangerous chemicals. Part 24: Vitro test of cell immunologic function
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| SN/T 2497.24-2010 | English | 139 |
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Test method of import and export dangerous chemicals. Part 24: Vitro test of cell immunologic function
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SN/T 2497.24-2010
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Basic data | Standard ID | SN/T 2497.24-2010 (SN/T2497.24-2010) | | Description (Translated English) | Test method of import and export dangerous chemicals. Part 24: Vitro test of cell immunologic function | | Sector / Industry | Commodity Inspection Standard (Recommended) | | Classification of Chinese Standard | G09 | | Classification of International Standard | 13.300 | | Word Count Estimation | 5,579 | | Date of Issue | 2010-03-02 | | Date of Implementation | 2010-09-16 | | Adopted Standard | Toxicology experimental methods and techniques in the immune toxicology research methods, MOD | | Regulation (derived from) | National Quality Inspection (2010) 98; industry standard filing Notice 2010 No. 5 (No. 125 overall) | | Issuing agency(ies) | General Administration of Customs | | Summary | This standard specifies the import and export of dangerous chemicals in vitro cellular immune function test principle, test methods and test results. This standard is applicable to the detection of import and export of dangerous chemicals in vitro immune toxicity testing. | SN/T 2497.24-2010: Test method of import and export dangerous chemicals. Part 24: Vitro test of cell immunologic function
---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
Test method of import and export dangerous chemicals.Part 24. Vitro test of cell immunologic function
Exit inspection and quarantine industry standard book People's Republic of China
Import and export of dangerous chemicals - Test methods
Part 24. Detection of cellular immune function in vitro
Issued on. 2010-03-02
2010-09-16 implementation
People's Republic of China
The State Administration of Quality Supervision, Inspection and Quarantine released
Foreword
SN/T 2497 "Test Methods for Chemical Safety & Exports dangerous" series of standards is divided into 29 parts.
--- Part 1. mammalian liver cells in vivo DNA synthesis (UDS) test;
--- Part 2. plaque-forming cells (PFC) test;
--- Part 3. Large Zhuan reproduction test;
--- Part 4. Saccharomyces cerevisiae mitotic recombination test;
--- Part 5. testicular cells UDS test;
--- Part 6. mammalian cells in vitro sister chromatid exchange test;
--- Part 7. mouse ear swelling test;
Part --- Article 8. fossa lymph node assay;
--- Part 9. Determination of serum hemolysin test;
Part --- Article 10. T lymphocyte proliferation assay test;
--- Part 11. germline mutation assay;
--- Part 12. single cell gel electrophoresis test;
--- Part 13. fluorescence in situ hybridization;
Part --- Article 14. SDS- polyacrylamide gel electrophoresis;
--- Part 15. PCR-SSCP experiment;
Part --- Article 16. Western-Blot test;
--- Part 17. Behavior of mammalian toxicology tests;
--- Part 18. Detection of DNA adducts;
--- Part 19. NorthernBlot experiment;
--- Part 20. Bradford protein assay;
--- Part 21. agarose gel electrophoresis;
--- Part 22. Tm value determination method of DNA;
--- Part 23. organelle separation experimental methods;
--- Part 24. cellular immunity in vitro assay method;
--- Part 25. humoral immune function tests;
--- Part 26. macrophage function tests;
--- Part 27. Flow cytometry of apoptosis;
--- Part 28. Application shuttle plasmid mutations Inspection;
--- Part 29. Biochemical Oxygen Demand (BOD) was measured.
This part is Part 24 SN/T 2497 series of standards.
The partial modification of the use of "Toxicology test methods and techniques" immune toxicology research method, technical content and the method described above
Exactly the same, in a standard text format according to GB/T 1.1 made editorial changes.
This section proposed and managed by the National Certification and Accreditation Administration Committee.
This section is responsible for drafting unit. People's Republic of China Tianjin Entry-Exit-Entry Inspection and Hunan People's Republic of China
Quarantine Bureau.
The main drafters of this section. Lu Gang, Wangli Bing, Chi Yu Feng, Li Xue Yang, Xiongzhong Jiang, Zhao good Lippo.
The first part of the Department of Inspection and Quarantine issued by industry standards.
Import and export of dangerous chemicals - Test methods
Part 24. Detection of cellular immune function in vitro
1 Scope
SN/T 2497 provisions of this part of the test principle and export of dangerous chemicals in the immune function test in vitro tests, test
Test methods and test results.
This section applies to in vitro testing for immunotoxicity testing and export of dangerous chemicals in cells.
2 test principle
Human peripheral blood T lymphocytes have receptors sheep erythrocytes (E receptor), that CD2 molecules, and B lymphocytes are not.
Thus in vitro, when mixed with sheep red blood cells, the T cells can be formed as the center, surrounded by sheep red blood cells
Like garlands of roses. This test can be used to detect the number and proportion of peripheral blood T lymphocytes.
Test Method 3
3.1 Reagents
3.1.1 lymphocyte separation medium, heparin, calcium, magnesium, Hank's solution, pH value of 7.2 ~ 7.6, Alsever solution, 0.8% glutaraldehyde,
0.2% cold Meilan.
3.1.2 sheep red blood cells
Fresh sheep blood was washed with a solution to save Alsever, formulation of 1% sheep red blood cell suspension.
3.1.3 Absorption inactivated fetal bovine serum
After inactivated fetal bovine serum after 56 ℃ 30min per ml of fetal calf serum was added to the washed sheep red blood hematocrit 0.1mL, mixing,
37 ℃ for 30min, then 2000r/min centrifugal 20min, supernatant at 4 ℃.
3.2 Test procedure
3.2.1 take anti-clotting heparin 3mL, plus 3mLHank's solution to fold diluted blood was added slowly to the solution of lymphocyte separation along the wall
On, 2000r/min centrifugal 20min.
3.2.2 Carefully draw isolated white liquid interface layer of mononuclear cells (containing lymphocytes), washed with Hank's solution twice, to adjust the cell
At a concentration of 2 × 106 cells/mL spare.
3.2.3 lymphocyte suspension was added 0.1mL Fahrenheit in a new tube, the absorption inactivated fetal bovine serum 0.1mL, 1% sheep red blood cells
Suspension 0.1mL mix, 37 ℃ incubated for 5min, then 500r/min centrifugal 5min. The tubes were placed in 4 ℃ refrigerator, at least 2h.
3.2.4 Remove the test tube, the supernatant was discarded part, gently rotate the tube Fahrenheit cells mix, then along the wall of one drop of 0.8% glutaraldehyde,
Placed 4 ℃ refrigerator 20min.
3.2.5 gently draw sheet 1 drops, 1 drop of 0.2% plus cold methylene blue dye 5min, cover glass, high magnification observation rosette formation
Happening.
4 results
Where the adsorption lymphocytes around 3 or more persons shall E- SRBC rosette forming positive lymphocyte count of 200,
Percentages E- rosette formation, see equation (1).
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