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Detection method for folic acid concentration for maintaining human lymphocyte cell genome stability in vitro

A technology for genome stability and lymphocytes, applied in the field of in vitro maintenance of human lymphocyte genome stability folic acid concentration detection, can solve problems such as chromosome breakage, centromere heterochromatin condensation level, gene expression changes, etc., to achieve maintenance The effect of stability

Inactive Publication Date: 2017-06-13
YUNNAN NORMAL UNIV
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Problems solved by technology

If the intake of folic acid is insufficient, the synthesis of dTMP will be hindered, and the excessive accumulation of dUMP will be incorporated into DNA, which will affect the subsequent DNA replication and repair process, and induce genetic structures such as base mismatch, gene mutation, DNA single and double strand breaks, and chromosome breaks. At the same time, it causes the main methyl donor-SAM reserve of most of the methylation reactions in the body to change, which changes the degree of DNA methylation in the whole genome and the methylation pattern of specific DNA sites, resulting in changes in gene expression, Genotoxic and epigenotoxic events such as decreased levels of centromere heterochromatin condensation occur

Method used

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Embodiment

[0024] A method for detecting the concentration of folic acid for maintaining genome stability of human lymphocytes in vitro, the steps comprising:

[0025] (1) Design culture medium: according to the folic acid content in human blood, set the folic acid concentration of 30nM, 60nM, 120nM, 240nM and configure it into 10ml, and the other components and concentrations of the medium are consistent with RPMI-1640 medium (Table 1);

[0026] Table 1 Preparation of RPMI-1640 medium without folic acid

[0027] Preparation volume 100(ml) RPMI 1640 culture medium (no folic acid) 90 Dialyzed calf serum (ml) 8 Final concentration of L-glutamine stock solution (200mM): 0.3mg / ml 1 Double antibody stock solution (100000U / ml) final concentration: 100U / ml 0.1 Final concentration of IL-2 stock solution (10000U / ml): 10U / ml 0.1

[0028] (2) Lymphocyte separation:

[0029] ①Take 5ml of blood from the vein into a clean sterile test tube with heparin, add...

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Abstract

The invention relates to the technical field of molecular biology and in particular relates to a detection method for folic acid concentration for maintaining human lymphocyte cell genome stability in vitro. The detection method comprises the following steps: designing a culture medium; separating lymphocyte cells; culturing the lymphocyte cells; collecting the cells and preparing a sheet; analyzing the sheet; carrying out statistic analysis. The human peripheral blood lymphocyte cells are cultured in vitro and different folic acid concentrations are set; the optimal concentration for maintaining the human lymphocyte cell genome stability in vitro is detected through CBMN (Cytokinesis Block Micronucleus) Cyt and the measured optimal concentration for maintaining the human lymphocyte cell genome stability in vitro is 120nmol / L; a result is closer to a living body and important evidences are provided for nutrition intervention of individuals and effective maintenance of the individual genome stability.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a method for detecting the concentration of folic acid for maintaining the genome stability of human lymphocytes in vitro. Background technique [0002] The genome carries all the biological information necessary for the formation and maintenance of human life, enabling humans to not only carry out various life activities normally in the long-term evolution process, but also to stably inherit various traits adapted to the environment to future generations. The relative stability of the human genome is taken as a premise. With the in-depth study of epigenetic phenomena and the continuous expansion of genome information knowledge, people have discovered that in addition to gene mutations, chromosomal aberrations, gene dosage changes caused by aneuploidy, and chromosome breakage-fusion-bridge (breakage-fusion-bridge) , BFB) cycles lead to gene amplification and other geno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02
CPCG01N33/5008G01N33/5026G01N33/505G01N33/5052
Inventor 汪旭王晗倪娟薛京伦
Owner YUNNAN NORMAL UNIV
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