CN106701683A - Establishing method for cell model used for researching drug absorption under plateau anaerobic condition - Google Patents

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

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CN106701683A
CN106701683A CN201611113372.0A CN201611113372A CN106701683A CN 106701683 A CN106701683 A CN 106701683A CN 201611113372 A CN201611113372 A CN 201611113372A CN 106701683 A CN106701683 A CN 106701683A
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王荣
赵安鹏
贾正平
李文斌
罗冰峰
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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

Method for building up for studying the cell model of drug absorption in the case of high altitude anoxia
Technical field
The present invention relates to Pharmacokinetics research field, medicine is inhaled in the case of more particularly, to studying high altitude anoxia The method for building up of the cell model of receipts.
Background technology
China's plateau territory total area is occupied first of the world, is possessed large number of plateau permanent resident population and is migrated population, this Outward, every year because the reasons such as tourism, work, military mission enter the number of highlands more than 10,000,000 person-times by plains region. With the increasingly increase of plateau crowd, plateau drug safety problem has turned into the potential threat of plateau population health.
Altitude makes body produce a series of physiologicals to change, partly for pathologic changes, these change influence medicines Absorption, distribution, metabolism and excretion in vivo, cause pharmacokinetic profile to change.However, the use of current medicine Method consumption does not consider the particularity on plateau, is that plateau drug safety hides some dangers for.
At present, the mammals such as rat, rabbit are focused primarily upon for the research mode of plateau pharmacokinetics Plateau site test and boiler-plate are tested.Zhang Juanhong etc. proved using the radical 4010 meters of plateau site tests of rat, Propranolol Drug-time curve under area（AUC）, average residence time（MRT）, half-life period（t1/2）, Cmax（Cmax）Significantly increase, CL, apparent volume of distribution (Vd) are significantly reduced；Kerdi etc. is thanked with low-pressure oxygen cabin simulated hypoxia Environmental Studies for rabbit Enzyme, finds the activity and expression generation significant changes of P450 enzymes after acute anoxia 48h.Also there are the human trial of minority, Li Xiangyang There is significant change in absorption and metabolism radical and occupy long in the healthy volunteer's body of 3800m plateaus Deng discovery Sulfamethoxazole, It is mainly shown as CL reductions, t1/2Extension.Plateau site test expends a large amount of manpower and materials, and boiler-plate experiment then needs costliness Large-scale instrument, and do not study the study in vitro of plateau pharmacokinetics clearly at present.
Caco-2 cell models are isolated from human colon cancer cells first in 20 century 70s, cultivate ripe Caco-2 Cell can form the monolayer organization with human small intestine's epithelial cell identical cell polarity and densification, and form and function are small with human body Enterocyte is very much like, has been widely used in Pharmacokinetics research, the external sieve as Oral drug absorption Modeling type.However, having no the experimental technique using drug transport in the case of Caco-2 scale-model investigation high altitude anoxias at present.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of simple and easy to apply, economic science for studying high altitude anoxia In the case of drug absorption cell model method for building up.
It is of the present invention for studying the cell model of drug absorption in the case of high altitude anoxia to solve the above problems Method for building up, comprises the following steps：
By Caco-2 cells use DMEM in high glucose nutrient solution in volumetric concentration for 5% CO2Cultivated in incubator, liquid is changed every other day Once, to passing on next time；
Using the step (1) gained Caco-2 cells on the filter membrane of 12 orifice plate Transwell cells build Caco-2 it is thin Born of the same parents' single-layer model；
(3) the culture medium in the Transwell cells is discarded, the Caco-2 cell monolayers model is washed 2 ~ 4 times using HBSS；
(4) determine the step (3) gained Caco-2 cell monolayer models cross-film resistance value, when cross-film resistance value be more than 400 Then it is qualified cell Caco-2 cell monolayer models during Ω；
(5) 12 ~ 48h of anoxic culture is carried out to the qualified cell Caco-2 cell monolayers model；
(6) the HBSS in the Transwell cells is discarded, new HBSS is added, put on 37 DEG C of distant beds of level, to the step (5) 30 ~ 60min is washed with the speed of 50rpm through the Caco-2 cell monolayers model of anoxic culture；
(7) determine the step (6) gained Caco-2 cell monolayer models cross-film resistance value, fluorescein apparent permeability coefficients, When cross-film resistance value is less than 1 × 10 more than 400 Ω, fluorescein apparent permeability coefficients-6Model evaluation can be completed during cm/s.
The step (1) middle Caco-2 cells be the 40th ~ 50 instead of between cell.
The step (1) middle DMEM in high glucose nutrient solution refer to by hyclone 100mL, 100 × nonessential amino acid 10mL, HEPES 5.96g, M Glus 0.06g, the unit of penicillin 80000, streptomysin 100mg are dissolved in 890mL DMEM sugar trainings high Support the nutrient solution of gained in base.
(1) middle condition of culture refers to that temperature is 36.5 ~ 37.5 DEG C to the step, and humidity is 85 ~ 95%.
(2) the construction method of middle Caco-2 cell monolayers model refers to be incited somebody to action using DMEM in high glucose nutrient solution first to the step The step (1) gained Caco-2 cells be made 2 × 106The suspension of individual/mL；Then 1 × 10 is pressed5Individual/cm2Caco-2 is thin for inoculation In on the filter membrane of the 12 orifice plate Transwell cells, culture is obtained final product born of the same parents for 21 ~ 28 days；Liquid, first 2 weeks are changed after wherein 8 ~ 16h of inoculation Liquid is changed every other day, changes liquid daily afterwards.
(3) (6) middle HBSS each means and for glucose 4.5g, HEPES 5.96g to be dissolved in 1mL the step with the step Hank ' s liquid, and adjust pH value to 7.2 ~ 7.4 with the HCl solution that mass concentration is 4% or NaOH solution that mass concentration is 4% Solution.
(5) middle anoxic culture refers to that the qualified cell Caco-2 cell monolayer models are put into volume is dense to the step It is 1 ~ 5%O to spend2And volumetric concentration is 5%CO2Three gas incubators in cultivate 12 ~ 48h.
(7) the assay method of fluorescein apparent permeability coefficients refers to discard to swing first to wash 30 ~ 60min's to the step The HBSS of the small indoor and outdoors of Transwell；Then Transwell cells are put into 12 new orifice plates, and small indoor adds 1mg/mL fluorescein solution 0.5mL, outside adds 1.5mL HBSS；Finally, shaked with the convolution of 50rpm rate levels in 37 DEG C, Outside HBSS absorption values at 460nm are determined using ELIASA after 2h, and fluorescein content is calculated with standard curve, as the following formula Calculate apparent permeability coefficients：Paap=C·V/（T·A·C0）
Wherein：Papp is fluorescein apparent permeability coefficients；C is to measure fluorescein concentration, unit %；V is reception tank volume, unit cm3；T is transhipment time, unit s；A is Transwell filter membrane areas, unit cm2；C0It is supply pool initial concentration, unit %.
The present invention has advantages below compared with prior art：
The present invention, by anoxic culture, is established Pharmaceutical sausage is inhaled suitable for research high altitude anoxia based on Caco-2 models The external pharmacokinetics cell model of influence is received, simple to operate, economic science is capable of achieving high throughput assay, to plateau Crowd's personalized medicine, the rational use of medicines has larger facilitation.
Brief description of the drawings
Specific embodiment of the invention is described in further detail below in conjunction with the accompanying drawings.
Fig. 1 is schematic diagram of the invention.
Specific embodiment
Embodiment 1 is used to study the method for building up of the cell model of drug absorption in the case of high altitude anoxia, including following step Suddenly：
(1) use DMEM in high glucose nutrient solution in temperature for CO that 37 DEG C, humidity are that 90%, volumetric concentration is 5% in Caco-2 cells2 Cultivated in incubator, liquid is changed every other day once, to passing on next time.
Wherein：Caco-2 cells be the 40th ~ 50 instead of between cell.
DMEM in high glucose nutrient solution refers to by hyclone 100mL, 100 × nonessential amino acid 10mL, HEPES 5.96g, M Glu 0.06g, the unit of penicillin 80000, streptomysin 100mg are dissolved in the training of gained in 890mL DMEM high glucose mediums Nutrient solution.
Using step (1) gained Caco-2 cells on the filter membrane of 12 orifice plate Transwell cells build Caco-2 it is thin Born of the same parents' single-layer model.
Wherein：The construction method of Caco-2 cell monolayer models refers to by step (1) institute first using DMEM in high glucose nutrient solution The Caco-2 cells for obtaining are made 2 × 106The suspension of individual/mL；Then 1 × 10 is pressed5Individual/cm2Inoculation Caco-2 cells are in 12 orifice plates On the filter membrane of Transwell cells, 8 holes are inoculated with altogether；Culture is obtained final product for 21 days；Liquid is changed after wherein inoculation 8h, is changed every other day within first 2 weeks Liquid, changes liquid daily afterwards.
(3) the culture medium in Transwell cells is discarded, Caco-2 cell monolayers model is washed 2 times using HBSS.
Wherein：HBSS each means and for glucose 4.5g, HEPES 5.96g to be dissolved in 1mL hank ' s liquid, and uses mass concentration For 4% HCl solution or mass concentration be 4% NaOH solution adjust pH value to 7.2 ~ 7.4 solution.
Determination step (3) gained Caco-2 cell monolayer models cross-film resistance value, when cross-film resistance value be more than 400 Then it is qualified cell Caco-2 cell monolayer models during Ω.
(5) anoxic culture 12h is carried out to qualified cell Caco-2 cell monolayers model, cell Caco-2 that will be qualified Cell monolayer model is put into volumetric concentration for 2%O2And volumetric concentration is 5%CO2Three gas incubators in cultivate 12h.
(6) the HBSS in Transwell cells is discarded, new HBSS is added, put on 37 DEG C of distant beds of level, (5) step is passed through The Caco-2 cell monolayers model of anoxic culture washes 30min with the speed of 50rpm.
Wherein HBSS with step (3).
Determination step (6) gained Caco-2 cell monolayer models cross-film resistance value, fluorescein apparent permeability coefficients, When cross-film resistance value is less than 1 × 10 more than 400 Ω, fluorescein apparent permeability coefficients-6Model evaluation can be completed during cm/s.
The assay method of fluorescein apparent permeability coefficients refers to discard to swing the small interiors of Transwell for washing 30 ~ 60min first Outer HBSS；Then Transwell cells are put into 12 new orifice plates, and small indoor adds 1mg/mL fluorescein solution 0.5mL, outside adds 1.5mL HBSS；Finally, shaked with the convolution of 50rpm rate levels in 37 DEG C, surveyed using ELIASA after 2h Fixed outside HBSS absorption values at 460nm, and fluorescein content is calculated with standard curve, it is calculated as follows apparent infiltration system Number：Paap=C·V/（T·A·C0）
Wherein：Papp is fluorescein apparent permeability coefficients；C is to measure fluorescein concentration, unit %；V is reception tank volume, unit cm3；T is transhipment time, unit s；A is Transwell filter membrane areas, unit cm2；C0It is supply pool initial concentration, unit %.
Embodiment 2 is used to study the method for building up of the cell model of drug absorption in the case of high altitude anoxia, including following step Suddenly：
(1) use DMEM in high glucose nutrient solution in temperature for 36.5 DEG C, humidity are that 85%, volumetric concentration is 5% in Caco-2 cells CO2Cultivated in incubator, liquid is changed every other day once, to passing on next time.
Wherein Caco-2 cells, DMEM in high glucose nutrient solution are with embodiment 1.
Using step (1) gained Caco-2 cells on the filter membrane of 12 orifice plate Transwell cells build Caco-2 it is thin Born of the same parents' single-layer model.
Wherein：The construction method of Caco-2 cell monolayer models refers to by step (1) institute first using DMEM in high glucose nutrient solution The Caco-2 cells for obtaining are made 2 × 106The suspension of individual/mL；Then 1 × 10 is pressed5Individual/cm2Inoculation Caco-2 cells are in 12 orifice plates On the filter membrane of Transwell cells, 12 holes are inoculated with altogether；Culture is obtained final product for 23 days；Liquid is changed after wherein inoculation 12h, is changed every other day within first 2 weeks Liquid, changes liquid daily afterwards.
(3) the culture medium in Transwell cells is discarded, Caco-2 cell monolayers model is washed 3 times using HBSS.
Wherein HBSS is with embodiment 1.
Determination step (3) gained Caco-2 cell monolayer models cross-film resistance value, when cross-film resistance value be more than 400 Then it is qualified cell Caco-2 cell monolayer models during Ω.
(5) anoxic culture 24h is carried out to qualified cell Caco-2 cell monolayers model, cell Caco-2 that will be qualified Cell monolayer model is put into volumetric concentration for 2%O2And volumetric concentration is 5%CO2Three gas incubators in cultivate 24h.
(6) the HBSS in Transwell cells is discarded, new HBSS is added, put on 37 DEG C of distant beds of level, (5) step is passed through The Caco-2 cell monolayers model of anoxic culture washes 40min with the speed of 50rpm.
Wherein HBSS with step (3).
Determination step (6) gained Caco-2 cell monolayer models cross-film resistance value, fluorescein apparent permeability coefficients, When cross-film resistance value is less than 1 × 10 more than 400 Ω, fluorescein apparent permeability coefficients-6Model evaluation can be completed during cm/s.
Wherein the assay method of fluorescein apparent permeability coefficients is with embodiment 1.
Embodiment 3 is used to study the method for building up of the cell model of drug absorption in the case of high altitude anoxia, including following step Suddenly：
(1) use DMEM in high glucose nutrient solution in temperature for 37.5 DEG C, humidity are that 95%, volumetric concentration is 5% in Caco-2 cells CO2Cultivated in incubator, liquid is changed every other day once, to passing on next time.
Wherein Caco-2 cells, DMEM in high glucose nutrient solution are with embodiment 1.
Using step (1) gained Caco-2 cells on the filter membrane of 12 orifice plate Transwell cells build Caco-2 it is thin Born of the same parents' single-layer model.
Wherein：The construction method of Caco-2 cell monolayer models refers to by step (1) institute first using DMEM in high glucose nutrient solution The Caco-2 cells for obtaining are made 2 × 106The suspension of individual/mL；Then 1 × 10 is pressed5Individual/cm2Inoculation Caco-2 cells are in 12 orifice plates On the filter membrane of Transwell cells, 6 holes are inoculated with altogether；Culture is obtained final product for 25 days；Liquid is changed after wherein inoculation 16h, is changed every other day within first 2 weeks Liquid, changes liquid daily afterwards.
(3) the culture medium in Transwell cells is discarded, Caco-2 cell monolayers model is washed 4 times using HBSS.
Wherein HBSS is with embodiment 1.
Determination step (3) gained Caco-2 cell monolayer models cross-film resistance value, when cross-film resistance value be more than 400 Then it is qualified cell Caco-2 cell monolayer models during Ω.
(5) anoxic culture 48h is carried out to qualified cell Caco-2 cell monolayers model, cell Caco-2 that will be qualified Cell monolayer model is put into volumetric concentration for 1%O2And volumetric concentration is 5%CO2Three gas incubators in cultivate 48h.
(6) the HBSS in Transwell cells is discarded, new HBSS is added, put on 37 DEG C of distant beds of level, (5) step is passed through The Caco-2 cell monolayers model of anoxic culture washes 50min with the speed of 50rpm.
Wherein HBSS with step (3).
Determination step (6) gained Caco-2 cell monolayer models cross-film resistance value, fluorescein apparent permeability coefficients, When cross-film resistance value is less than 1 × 10 more than 400 Ω, fluorescein apparent permeability coefficients-6Model evaluation can be completed during cm/s.
Wherein the assay method of fluorescein apparent permeability coefficients is with embodiment 1.
Embodiment 4 is used to study the method for building up of the cell model of drug absorption in the case of high altitude anoxia, including following step Suddenly：
(1) use DMEM in high glucose nutrient solution in temperature for CO that 37 DEG C, humidity are that 90%, volumetric concentration is 5% in Caco-2 cells2 Cultivated in incubator, liquid is changed every other day once, to passing on next time.
Wherein Caco-2 cells, DMEM in high glucose nutrient solution are with embodiment 1.
Using step (1) gained Caco-2 cells on the filter membrane of 12 orifice plate Transwell cells build Caco-2 it is thin Born of the same parents' single-layer model.
Wherein：The construction method of Caco-2 cell monolayer models refers to by step (1) institute first using DMEM in high glucose nutrient solution The Caco-2 cells for obtaining are made 2 × 106The suspension of individual/mL；Then 1 × 10 is pressed5Individual/cm2Inoculation Caco-2 cells are in 12 orifice plates On the filter membrane of Transwell cells, 6 holes are inoculated with altogether；Culture is obtained final product for 28 days；Liquid is changed after wherein inoculation 14h, is changed every other day within first 2 weeks Liquid, changes liquid daily afterwards.
(3) the culture medium in Transwell cells is discarded, Caco-2 cell monolayers model is washed 2 times using HBSS.
Wherein HBSS is with embodiment 1.
Determination step (3) gained Caco-2 cell monolayer models cross-film resistance value, when cross-film resistance value be more than 400 Then it is qualified cell Caco-2 cell monolayer models during Ω.
(5) anoxic culture 36h is carried out to qualified cell Caco-2 cell monolayers model, cell Caco-2 that will be qualified Cell monolayer model is put into volumetric concentration for 5%O2And volumetric concentration is 5%CO2Three gas incubators in cultivate 36h.
(6) the HBSS in Transwell cells is discarded, new HBSS is added, put on 37 DEG C of distant beds of level, (5) step is passed through The Caco-2 cell monolayers model of anoxic culture washes 60min with the speed of 50rpm.
Wherein HBSS with step (3).
Determination step (6) gained Caco-2 cell monolayer models cross-film resistance value, fluorescein apparent permeability coefficients, When cross-film resistance value is less than 1 × 10 more than 400 Ω, fluorescein apparent permeability coefficients-6Model evaluation can be completed during cm/s.
Wherein the assay method of fluorescein apparent permeability coefficients is with embodiment 1.

Claims (8)

1. it is used to study the method for building up of the cell model of drug absorption in the case of high altitude anoxia, comprises the following steps：
By Caco-2 cells use DMEM in high glucose nutrient solution in volumetric concentration for 5% CO2Cultivated in incubator, liquid one is changed every other day It is secondary, to passing on next time；
Using the step (1) gained Caco-2 cells on the filter membrane of 12 orifice plate Transwell cells build Caco-2 it is thin Born of the same parents' single-layer model；
(3) the culture medium in the Transwell cells is discarded, the Caco-2 cell monolayers model is washed 2 ~ 4 times using HBSS；
(4) determine the step (3) gained Caco-2 cell monolayer models cross-film resistance value, when cross-film resistance value be more than 400 Then it is qualified cell Caco-2 cell monolayer models during Ω；
(5) 12 ~ 48h of anoxic culture is carried out to the qualified cell Caco-2 cell monolayers model；
(6) the HBSS in the Transwell cells is discarded, new HBSS is added, put on 37 DEG C of distant beds of level, to the step (5) 30 ~ 60min is washed with the speed of 50rpm through the Caco-2 cell monolayers model of anoxic culture；
(7) determine the step (6) gained Caco-2 cell monolayer models cross-film resistance value, fluorescein apparent permeability coefficients, When cross-film resistance value is less than 1 × 10 more than 400 Ω, fluorescein apparent permeability coefficients-6Model evaluation can be completed during cm/s.
2. the as claimed in claim 1 method for building up for being used to studying the cell model of drug absorption in the case of high altitude anoxia, its It is characterised by：The step (1) middle Caco-2 cells be the 40th ~ 50 instead of between cell.
3. the as claimed in claim 1 method for building up for being used to studying the cell model of drug absorption in the case of high altitude anoxia, its It is characterised by：The step (1) middle DMEM in high glucose nutrient solution refer to by hyclone 100mL, 100 × nonessential amino acid 10mL, HEPES 5.96g, M Glus 0.06g, the unit of penicillin 80000, streptomysin 100mg are dissolved in 890mL DMEM sugar trainings high Support the nutrient solution of gained in base.
4. the as claimed in claim 1 method for building up for being used to studying the cell model of drug absorption in the case of high altitude anoxia, its It is characterised by：(1) middle condition of culture refers to that temperature is 36.5 ~ 37.5 DEG C to the step, and humidity is 85 ~ 95%.
5. the as claimed in claim 1 method for building up for being used to studying the cell model of drug absorption in the case of high altitude anoxia, its It is characterised by：(2) the construction method of middle Caco-2 cell monolayers model refers to be incited somebody to action using DMEM in high glucose nutrient solution first to the step The step (1) gained Caco-2 cells be made 2 × 106The suspension of individual/mL；Then 1 × 10 is pressed5Individual/cm2Caco-2 is thin for inoculation In on the filter membrane of the 12 orifice plate Transwell cells, culture is obtained final product born of the same parents for 21 ~ 28 days；Liquid, first 2 weeks are changed after wherein 8 ~ 16h of inoculation Liquid is changed every other day, changes liquid daily afterwards.
6. the as claimed in claim 1 method for building up for being used to studying the cell model of drug absorption in the case of high altitude anoxia, its It is characterised by：(3) (6) middle HBSS each means and for glucose 4.5g, HEPES 5.96g to be dissolved in 1mL the step with the step Hank ' s liquid, and adjust pH value to 7.2 ~ 7.4 with the HCl solution that mass concentration is 4% or NaOH solution that mass concentration is 4% Solution.
7. the as claimed in claim 1 method for building up for being used to studying the cell model of drug absorption in the case of high altitude anoxia, its It is characterised by：(5) middle anoxic culture refers to that the qualified cell Caco-2 cell monolayer models are put into volume is dense to the step It is 1 ~ 5%O to spend2And volumetric concentration is 5%CO2Three gas incubators in cultivate 12 ~ 48h.
8. the as claimed in claim 1 method for building up for being used to studying the cell model of drug absorption in the case of high altitude anoxia, its It is characterised by：(7) the assay method of fluorescein apparent permeability coefficients refers to discard to swing first to wash 30 ~ 60min's to the step The HBSS of the small indoor and outdoors of Transwell；Then Transwell cells are put into 12 new orifice plates, and small indoor adds 1mg/mL fluorescein solution 0.5mL, outside adds 1.5mL HBSS；Finally, shaked with the convolution of 50rpm rate levels in 37 DEG C, Outside HBSS absorption values at 460nm are determined using ELIASA after 2h, and fluorescein content is calculated with standard curve, as the following formula Calculate apparent permeability coefficients：Paap=C·V/（T·A·C0）
Wherein：Papp is fluorescein apparent permeability coefficients；C is to measure fluorescein concentration, unit %；V is reception tank volume, unit cm3；T is transhipment time, unit s；A is Transwell filter membrane areas, unit cm2；C0It is supply pool initial concentration, unit %.
CN201611113372.0A 2016-12-07 2016-12-07 Establishing method for cell model used for researching drug absorption under plateau anaerobic condition Pending CN106701683A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107312750A (en) * 2017-08-02 2017-11-03 烟台大学 For the method for building up for the myocardial fibrosis cell model for studying drug absorption
CN111690591A (en) * 2020-06-19 2020-09-22 贵州中医药大学 Diuretic active drug cell screening model and establishment method and application thereof
CN113621575A (en) * 2021-08-17 2021-11-09 山东大学 Application of fucosan sulfate in cell model construction

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
王文生: "肠缺血再灌注条件下IFN-γ通过调控HIF-1α在肠上皮屏障功能损害中的作用及机制研究", 《中国博士学位论文全文数据库 医学卫生科技辑》 *
马美湖、黄晶: ""Caco-2 细胞模型快速培养法的建立与评价", 《现代食品科技》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107312750A (en) * 2017-08-02 2017-11-03 烟台大学 For the method for building up for the myocardial fibrosis cell model for studying drug absorption
CN111690591A (en) * 2020-06-19 2020-09-22 贵州中医药大学 Diuretic active drug cell screening model and establishment method and application thereof
CN111690591B (en) * 2020-06-19 2022-03-15 贵州中医药大学 Diuretic active drug cell screening model and establishment method and application thereof
CN113621575A (en) * 2021-08-17 2021-11-09 山东大学 Application of fucosan sulfate in cell model construction
CN113621575B (en) * 2021-08-17 2024-04-09 山东大学 Application of fucoidan sulfate in cell model construction

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Application publication date: 20170524