Establishing method for cell model used for researching drug absorption under plateau anaerobic condition

A technology of cell model and method establishment, applied in the field of pharmacokinetics research, to achieve the effects of simple operation, promotion of rational drug use, and economical science

Inactive Publication Date: 2017-05-24
王荣
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  • Description
  • Claims
  • Application Information

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However, there is currently no experimental method using th

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  • Establishing method for cell model used for researching drug absorption under plateau anaerobic condition

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Embodiment 1

[0027] Example 1 The method for establishing a cell model for studying drug absorption under plateau hypoxia conditions includes the following steps:

[0028] ⑴The Caco-2 cells were cultured in high-sugar DMEM medium at a temperature of 37°C, a humidity of 90%, and a volume concentration of 5% CO 2 Cultured in an incubator, the medium was changed every other day until the next passage.

[0029] Among them: Caco-2 cells are cells between the 40th and 50th passages.

[0030] High-sugar DMEM culture medium refers to the solution obtained by dissolving 100mL of fetal bovine serum, 10mL of 100×non-essential amino acids, 5.96g of HEPES, 0.06g of ML-glutamine, 80000 units of penicillin, and 100mg of streptomycin in 890mL of DMEM high-sugar medium culture fluid.

[0031] (2) Use the Caco-2 cells obtained in step (1) to construct a Caco-2 cell monolayer model on the filter membrane of the Transwell chamber of a 12-well plate.

[0032] Wherein: the construction method of Caco-2 cell ...

Embodiment 2

[0042] Example 2 The method for establishing a cell model for studying drug absorption under plateau hypoxia conditions includes the following steps:

[0043] ⑴The Caco-2 cells were cultured in high-sugar DMEM medium at a temperature of 36.5°C, a humidity of 85%, and a volume concentration of 5% CO 2 Cultured in an incubator, the medium was changed every other day until the next passage.

[0044] Wherein the Caco-2 cells and high-sugar DMEM culture fluid are the same as in Example 1.

[0045] (2) Use the Caco-2 cells obtained in step (1) to construct a Caco-2 cell monolayer model on the filter membrane of the Transwell chamber of a 12-well plate.

[0046] Wherein: the construction method of Caco-2 cell monolayer model refers to at first adopting high-sugar DMEM culture fluid to make the Caco-2 cell of step (1) gained 2 * 10 6 individual / mL suspension; then press 1×10 5 piece / cm 2 Inoculate Caco-2 cells on the filter membrane of a 12-well plate Transwell chamber, inoculate ...

Embodiment 3

[0055] Example 3 The method for establishing a cell model for studying drug absorption under plateau hypoxia conditions includes the following steps:

[0056] ⑴Caco-2 cells were cultured in high-sugar DMEM medium at a temperature of 37.5°C, a humidity of 95%, and a volume concentration of 5% CO 2 Cultured in an incubator, the medium was changed every other day until the next passage.

[0057] Wherein the Caco-2 cells and high-sugar DMEM culture fluid are the same as in Example 1.

[0058] (2) Use the Caco-2 cells obtained in step (1) to construct a Caco-2 cell monolayer model on the filter membrane of the Transwell chamber of a 12-well plate.

[0059] Wherein: the construction method of Caco-2 cell monolayer model refers to at first adopting high-sugar DMEM culture fluid to make the Caco-2 cell of step (1) gained 2 * 10 6 individual / mL suspension; then press 1×10 5 piece / cm 2 Inoculate Caco-2 cells on the filter membrane of a 12-well plate Transwell chamber, inoculate 6 we...

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Abstract

The invention relates to an establishing method for a cell model used for researching drug absorption under a plateau anaerobic condition. The establishing method comprises the following steps: (1) culturing Caco-2 cells in a high-sugar DMEM culture solution; (2) constructing a Caco-2 cell single-layer model on a filter membrane of a 12-pore plate Transwell cell by the Caco-2 cells obtained in the step (1); (3) washing the Caco-2 cell single-layer model by HBSS; (4) measuring transepithelial electrical resistance of the Caco-2 cell single-layer model obtained in the step (3), and determining the Caco-2 cell single-layer model as a qualified Caco-2 cell single-layer model when the transepithelial electrical resistance is greater than 400 ohm; (5) performing anaerobic culture on the qualified Caco-2 cell single-layer model; (6) cleaning the Caco-2 cell single-layer model subjected to the anaerobic culture in the step (5); and (7) measuring the transepithelial electrical resistance and an fluorescein apparent permeability coefficient of the Caco-2 cell single-layer model obtained in the step (6), and completing model evaluation. The establishing method is simple to operate, is economic and scientific, can realize high-throughput measurement, administrates drugs for highland people in an individualized mode, and has relatively great promotion effect on reasonable drug administration.

Description

technical field [0001] The invention relates to the research field of pharmacokinetics, in particular to a method for establishing a cell model for studying drug absorption under the condition of plateau hypoxia. Background technique [0002] The total land area of ​​my country's plateau ranks first in the world, and it has a large number of plateau permanent residents and immigrants. In addition, more than 10 million people enter the plateau area from the plain area every year due to tourism, work, military missions and other reasons. With the increasing number of plateau populations, the safety of medication at high altitudes has become a potential threat to the health of plateau populations. [0003] Plateau hypoxia causes the body to produce a series of physiological changes, some of which are pathological changes, which affect the absorption, distribution, metabolism and excretion of drugs in the body, resulting in changes in pharmacokinetic characteristics. However, t...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693
Inventor 王荣赵安鹏贾正平李文斌罗冰峰
Owner 王荣
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