New method for performing data correction by using cell metabolite relative content as cell number index

A technology of cell metabolism and metabolomics data, which is applied in the field of metabolomics data correction and rapid quenching of biochemical metabolism of in vitro cultured cells, to achieve efficient extraction, ensure accuracy and reliability, and simplify experimental operations

Inactive Publication Date: 2011-09-07
CHINA PHARM UNIV
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Problems solved by technology

[0008] The technical problem to be solved by the present invention is to improve the existing cell collection and metabolic quenching methods in the study of cell metabolomics, optimize the solvent extraction scheme, and use the identified small molecule metabolites as an indicator of cell number for the obtained data to solve a series of research problems in the study of cell metabolomics

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  • New method for performing data correction by using cell metabolite relative content as cell number index
  • New method for performing data correction by using cell metabolite relative content as cell number index
  • New method for performing data correction by using cell metabolite relative content as cell number index

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Embodiment

[0030] Cellular metabolites as indicators of cell number for correction of cellular metabolomics data - Madin-Darby canine kidney (MDCK) wild-type (MDCK-WT) and transfected human Mdr1 gene transfection Metabolomics Paradigm Study in Transfected (MDCK-MDR) Cells

[0031] 1. Experimental Protocol and Sample Collection

[0032] Serial volumes of cell suspension (A-F: 2.5, 2.0, 1.6, 1.2, 0.9, 0.6mL, MDCK-WT 600,000 / mL and MDCK-MDR 800,000 / mL) were seeded in six-well plates (n=4), and blanks were added The medium was such that the volume per space was 2.5 mL. The two cell lines were grown under the same culture conditions for about 3 days without changing the culture medium. The medium samples of the cells in each well were collected, and the cells were pretreated according to the above method, and 300 μL of ultrapure water was added to each well, and then frozen in a -70°C refrigerator for testing.

[0033] 2. Sample Processing

[0034] Cell sample: After repeated freezing and...

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Abstract

The invention discloses a new method for performing data correction by using cell metabolite relative content as cell number index. The new method is characterized by comprising the steps of: realizing the fast quenching of biochemical metabolism in cells by using a simple sample pre-processing method; efficiently extracting small molecule metabolites by an optimized extraction solvent; and roundly and half quantitatively measuring the small molecule compounds in a biological sample by a metabonomics method based on measuring technology like mass spectrum, nuclear magnetic resonance and the like so as to identify a quantitative index of symbolic metabolites in cells to be as the cell number, and regarding the quantitative index as a weight factor to perform data correction. Compared with the quantitative index of the prior cell number, the method is simple, fast and with good repeatability; additional experimental working and the error caused by the additional working are reduced; the accuracy and reliability of experiment result are improved; the difficulty for relatively quantifying the number of in vitro cultured cells is solved; a new displacing method is supplied for cell metabonomics and relevant researches; and the method can be widely used in the relevant researches in the field of cell molecular biology.

Description

technical field [0001] The present invention relates to a new method for using in vitro cultured cells to conduct metabolomics research related to diseases, drug efficacy, and drug toxicity. The content is used as a quantitative indicator of cell number and used in the method of metabolomics data correction. Background technique [0002] At present, the method of cell metabolomics research is still immature, especially the metabolomics research of adherent cells cultured in vitro is relatively lagging behind, and there are a series of problems, mainly in the following three aspects: [0003] 1. Cell Quantification Questions The difference in the proliferation rate of different cells, the same kind of cells are affected by the initial seeding density and different treatment methods (environmental or drug stimulation), which will lead to differences in the number of cells obtained by different cell lines or the same kind of cell lines grown in different states , it is neces...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N27/62G01N24/08G01N1/28C12Q1/02
Inventor 王广基阿基业曹蓓刘林生石建郑天李梦婕王新文赵春艳张凤逸
Owner CHINA PHARM UNIV
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